28. Arbeitstagung der
Anatomischen Gesellschaft
in Würzburg
28.09.2011 bis 30.09.2011
Vortrag 1
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:3
Titel: Does the spine apparatus contribute to synaptic transmission? - Functional
analysis of individual synapses using two-photon Ca2+ imaging combined with
glutamate uncaging in organotypic slice cultures
Autoren: Drakew A.(1),Tippmann A.(1),Maier U.(1),Frotscher M.(2),
Adressen:(1)Albert-Ludwigs-Universität Freiburg|Institut für Anatomie und
Zellbiologie, Abteilung Neuroanatomie|Freiburg|Deutschland;
email:alexander.drakew@anat.uni-freiburg.de; (2)Universitaetsklinikum HamburgEppendorf|Zentrum für Molekulare Neurobiologie Hamburg, Institut für
Strukturelle Neurobiologie|Hamburg|Deutschland
Abstract:
Glutamatergic synaptic excitation is often accompanied by a transient increase of
free cytoplasmic Ca2+ in dendritic spines. Postsynaptic Ca2+ stores potentially
contribute to these Ca2+ transients. It has been suggested that the spine
apparatus, which is a derivative of the endoplasmic reticulum, represents such a
Ca2+ store in dendritic spines. The large complex spines of hilar mossy cells and
CA3 pyramidal neurons contacted by the mossy fibers typically contain this
organelle. Mutant mice lacking the actin-binding protein Synaptopodin do not form
spine apparatuses in dendritic spines and show a reduced long-term potentiation
at the hippocampal CA3-CA1 synapse and an impaired spatial learning
performance [1].
To test the hypothesis that the spine apparatus contributes to Ca2+ kinetics in
synaptic transmission, we compared Ca2+ transients in large complex spines in
organotypic slice cultures of wild-type mice and Synaptopodin knockout mice.
The combination of the patch-clamp technique with two-photon Ca2+ imaging
and two-photon glutamate uncaging allowed us to correlate induced excitatory
postsynaptic events and the corresponding Ca2+ changes at individual synapses
of hilar mossy cells.
We found that in both genotypes the NMDA receptor is the main source of Ca2+
in response to single glutamate uncaging pulses. However, Ca2+ transients
induced by trains of uncaging pulses paired with backpropagating action
potentials had smaller amplitudes in tissue from Synaptopodin knockout mice.
This suggests that the spine apparatus is not necessary for basal synaptic
transmission but becomes involved during strong synaptic activation. [1] Deller et.
al. (2003) PNAS 100:10494-10499.
Supported by the DFG: SFB 780.
Kategorie: Vortrag
Vortrag 2
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: Plasticity-related gene 5 expression promotes spine formation in primary
neurons
Autoren: Bräuer A.(1),
Adressen:(1)Charitè Universitätsmedizin Berlin|Zell- und
Neurobiologie|Berlin|Germany; email:anja.braeuer@charite.de
Abstract:
The vast majority of excitatory synapses are made on the heads of dendritic
spines, highly dynamic structures. The shape of a neuron’s dendritic arbour
determines the set of axons with which it may form synaptic contacts, thus
establishing connectivity within neural circuits. Dynamic cytoskeleton remodelling
is an essential step during this process. The actin filament is belived to be the
basic structural foundation that is responsible for their shape. Membrane protein
may act through an intracellular signalling pathway, which ultimately converge on
the cytoskeleton. However, the molecular mechanisms involved in these steps
are not well understood. Here, we show firstly that overexpression of the integral
membrane protein Plasticity-related gene-5 (PRG-5), a member of the vertebrateand brain-specific PRG family, induce spines formation in young primary neurons,
at DIV 2. Mutagenesis experiments in PRG-5 show residual amino acids that are
important for the induction of spines. Furthermore, PRG-5 localizes to and
promotes mushroom spines formation in primary neurons at least DIV 10. siRNA
experiments, in mature primary neurons, confirm that endogenously level of PRG5 plays an importand role in spine formation.
Our results suggest that PRG-5 function is involved in spinogenesis process.
Kategorie: Vortrag
Vortrag 3
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:3
Titel: A role for synaptopodin in denervation-induced plasticity
Autoren: Vlachos A.(1),Ikenberg B.(1),Lenz M.(1),Bas Orth C.(1),Reifenberg
K.(2),Deller T.(1),
Adressen:(1)Goethe-University|Institute of Clinical
Neuroanatomy|Frankfurt|Germany; email:a.vlachos@med.uni-frankfurt.de;
(2)Johannes Gutenberg University|Central Laboratory Animal
Facility|Mainz|Germany
Abstract:
Neurons which are partially denervated after brain injury remodel their synaptic
connections and eventually achieve a new stable state. Although this lesioninduced synaptic reorganization has long been described, its dynamics and the
underlying molecular mechanisms are still insufficiently understood. Here, we
have investigated denervation-induced changes in excitatory synaptic strength of
granule cells after entorhinal denervation in organotypic slice cultures. Whole cell
patch clamp recordings revealed that partial deafferentation induces an increase
in excitatory synaptic strength, indicating that neurons upregulate their synaptic
strength in order to maintain net afferent drive. Since synaptopodin (SP) regulates
the accumulation of AMPA-R at spine postsynapses (Vlachos et al., 2009), we
speculated that the postsynaptic increase in excitatory synaptic strength requires
SP. Indeed, in entorhino-hippocampal slice cultures of SP-deficient animals
denervation-induced synaptic strengthening was not observed. By crossing the
SP-deficient mouse with a newly generated transgenic mouse strain which
expresses EGFP-tagged SP under the control of the Thy1.2. promoter, the ability
of dentate granule cells to increase their excitatory synaptic strength was
rescued. We conclude that SP is essential for the functional changes seen in
granule cells following entorhinal denervation. (Supported by GIF and DFG).
Kategorie: Vortrag
Vortrag 4
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:3
Titel: A sexual dimorphism in estrogen-induced synaptic plasticity
Autoren: Fester L.(1),Boehm J.(1),Jarry H.(2),Rune G.(1),
Adressen:(1)University Medical Center Hamburg-Eppendorf|Institute of
Neuroanatomy|Hamburg|Germany; email:lfester@uke.de; (2)University of
Goettingen|Institute of Experimental Endocrinology|Goettingen|Germany
Abstract:
In previous studies we showed, that changes of spine density during the estrus
cycle of females in the hippocampus depend on the regulation of local estrogen
synthesis in the hippocampus by GnRH. Furthermore we found, that synaptic
proteins, such as synaptophysin and spinophilin, are regulated by local
aromatase activity, the final enzyme in estradiol synthesis. GnRH regulates local
estradiol synthesis in the hippocampus via GnRH-receptors, which are most
prominent in the hippocampus as compared to the hypothalamus and the
neocortex. Since GnRH secretion is different in male and females we addressed
the question whether GnRH responsiveness in the hippocampus varies between
genders. First experiments indicate that GnRH stimulates estradiol synthesis in
females but inhibits estradiol synthesis in males. Accordingly, spine synapse
density is increased upon GnRH treatment in females. Since inhibition of estradiol
synthesis in males increases spine synapse density, GnRH treatment of
hippocampal cultures, that originated from males consistently resulted in
enhanced synapse density by downregulating estradiol synthesis. Our results
show a sexual dimorphism in estrogen-induced synaptic plasticity.
Kategorie: Vortrag
Vortrag 5
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:3
Titel: Aromatase activity is required for stabilizing the spine cytoskeleton
Autoren: Vierk R.(1),Zhou L.(1),Glassmeier G.(2),Brandt N.(1),Rune G.(1),
Adressen:(1)University Medical Center Hamburg-Eppendorf|Institute of
Neuroanatomy|Hamburg|Germany; email:r.vierk@uke.de; (2)University Medical
Center Hamburg-Eppendorf|Institute of Neurophysiology and
Pathophysiology|Hamburg|Germany
Abstract:
In this study we focussed on the functional significance of inhibition of
hippocampal estrogen synthesis, using long-term potentiation (LTP) as a cellular
model for memory. To this end, we reduced hippocampal estrogen synthesis in
hippocampal slice cultures and systemically in mice by using the aromatase
inhibitor letrozole. In acute slices of letrozole-treated mice as well as in
hippocampal slice cultures, letrozole treatment resulted in significant impairment
of LTP in response to theta burst stimulation of CA3-CA1 Schaffer collaterals.
The impairment was already seen after 6 hours of treatment and progressed
continuously. After 7 days of treatment, theta burst stimulation failed to induce
long-term potentiation. Consistently, letrozole decreased immunoreactivity of pcofilin in hippocampal cultures, since LTP is paralleled by an increase in F-actin
and by phosphorylation of cofilin, thereby stabilizing the spine cytoskeleton.
Impairment of LTP was followed by reduction of preferentially mushroom spines
on GFP-transfected dissociated neurons and final spine synapse loss in the
hippocampus of letrozole-treated animals. Our findings show that aromatase
activity is required for stabilizing the spine cytoskeleton which in turn, has been
shown to be a prerequisite for inducing LTP.
Kategorie: Vortrag
Vortrag 6
Rubrik: 8.Neuroregeneration/Neurodegeneration
Abstract Nr.:8
Titel: Synaptic connectivity in the pahenu2 mutant, a mouse model for
phenylketonuria
Autoren: Horling K.(1),Schlegel G.(2),Santer R.(3),Schumacher U.(1),Ullrich
K.(3),Rune G.(2),
Adressen:(1)University Medical Center Hamburg-Eppendorf|Departments of
Anatomy and Experimental Morphology|Hamburg|Germany;
email:k.horling@uke.uni-hamburg.de; (2)University Medical Center HamburgEppendorf|Department of Neuroanatomy|Hamburg|Germany; (3)University
Medical Center Hamburg-Eppendorf|Department of Pediatrics|Hamburg|Germany
Abstract:
Phenylketonuria (PKU) is an autosomal-recessive disorder with an incidence of
1:8000. An error in the genetic code of the hepatic enzyme phenylalanine
hydroxylase (PAH) causes increased levels of the amino acid phenylalanine in
blood and cerebrospinal fluid. In humans, phenylketonuria is associated with
mental retardation. The underlying mechanisms of this pathology are poorly
understood so far. Therefore, we used the PKU mouse model PAHenu2 with a
loss of function mutation of the PAH and similarly elevated brain phenylalanine
concentrations as in man for our experiments with respect to analyse
morphological changes in the brain. Most strikingly, in the hippocampus of these
mutants, stereological evaluation of spine synapse density revealed an increased
number of spine synapses in the PKU mouse as compared to the wildtype, both
in stratum radiatum of the CA1 and of the CA3 region. Immunoreactivity of
spinophilin and synaptopodin, postsynaptic proteins that are enriched in spines
and a marker for mature, mushroom shaped spines respectively, are significantly
increased in the mutants, as quantified by laser scanning microscopy and
subsequent image analysis. In contrast, SNAP25, which is involved in the release
of neurotransmitter at the presynaptic site, is markedly reduced in PAHenu2 mice.
In sum, synaptic connectivity is highly affected in response to elevated levels of
phenylalanine in the brain, which also may contribute to mental retardation in
humans.
Kategorie: Vortrag
Vortrag 7
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:3
Titel: Psa-ncam regulates synaptic targeting during development and modulates
regeneration after lesion
Autoren: Vogt J.(1),Schmitz D.(2),Heimrich B.(3),Nitsch R.(1),Horstkorte R.(4),
Adressen:(1)Universitätsmedizin Mainz|Institute of Microanatomy and
Neurobiology|Mainz|Deutschland; email:johannes.vogt@unimedizin-mainz.de;
(2)Charité – Universitätsmedizin Berlin|Neuroscience Research
Center|Berlin|Deutschland; (3)Albert-Ludwigs-University Freiburg|Institute for
Anatomy and Cell Biology|Freiburg|Deutschland; (4)Martin-Luther-Universität
Halle-Wittenberg|Institut für Physiologische Chemie|Halle/Saale|Deutschland
Abstract:
During development, axonal projections have a remarkable ability to innervate
correct dendritic subcompartiments of their target neurons. Altered axonal
targeting with formation of synapses on unappropriate neurons may result in
neurodevelopmental sequelae leading to psychiatric disorders. Here we show
that altering the expression of the polysialic acid moiety, which is a
developmentally regulated, posttranslational modification of the neural cell
adhesion molecule NCAM, critically affects correct circuit formation. Using a
chemically modified sialic acid precursor (N-propanoyl-D-mannosamine), we
inhibited the polysialyltransferase ST8SiaII, the principal enzyme involved in
polysialylation during development, at selected developmental timepoints. This
treatment altered NCAM polysialylation while NCAM-expression was not affected.
Altered polysialylation resulted in an aberrant mossy fiber projection which formed
glutamatergic terminals on pyramidal neurons of the CA1-region in organotypic
slice cultures and in vivo. Electrophysiological recordings revealed that the
ectopic terminals on CA1 pyramids were functional and displayed characteristics
of mossy fiber synapses. Moreover, ultrastructural examination indicated a
\"mossy fiber synapse”-like morphology. After mossy fiber tract lesions, activated,
ST8SiaII-expressing astrocytes formed a polysialic acid-positive glial scar
preventing further fiber outgrowth. Conversely, following ST8SiaII-inhibition
polysialic acid-synthesis was greatly diminished and astrocytes displayed a
quiescent, ramified morphology. Accordingly, astrocytes did not form a glial scar
allowing substantial reinnervation of the deafferentiated CA3 region by lesioned
mossy fiber-axons. We thus conclude that polysialic acid is essential for correct
outgrowth and synaptic targeting during mossy fiber development but may inhibit
regeneration by promoting astroglial scar formation after lesion.
Kategorie: Vortrag
Vortrag 8
Rubrik: 9.Peripheres und vegetatives Nervensystem
Abstract Nr.:9
Titel: Gdnf promotes enteric neuronal plasticity – implications for the
pathogenesis of diverticular disease.
Autoren: Böttner M.(1),Zorenkov D.(2),Bruch H.(3),Roblick U.(3),Egberts
J.(4),Schäfer K.(5),Wedel T.(1),
Adressen:(1)CAU Kiel|Anatomisches Insitut|Kiel|Schleswig-Holstein;
email:m.boettner@anat.uni-kiel.de; (2)Universitätsklinikum Schleswig-Holstein,
Campus Kiel|Klinik für Neurologie|Kiel|Schleswig-Holstein; (3)UKSH Campus
Lübeck|Chirurgie|Lübeck|Schleswig-Holstein; (4)UKSH Campus
Kiel|Chirurgie|Kiel|Schleswig-Holstein; (5)FH
Kaiserlslautern/Zweibrücken|Informatik und
Mikrosystemtechnik|Zweibrücken|Saarland
Abstract:
Background & aims:
Ablation of glial cell line-derived neurotrophic factor (GDNF) leads to intestinal
aganglionosis. We recently demonstrated that patients with diverticular disease
(DD) exhibit hypoganglionosis suggesting neurotrophic factor deprivation. Thus,
we screened mRNA expression of the GDNF system and markers of neuronal
plasticity in DD and examined the effect of GDNF on cultured enteric neurons.
Material & methods:
Human colonic samples (tunica muscularis) were assessed for mRNA expression
levels of the GDNF system (GDNF, RET, GFRa1/2) by a low-density PCR array.
To identify the tissue source of GDNF and its receptors, laser-microdissected
(LMD) samples of human myenteric ganglia and adjacent muscle layers were
analyzed separately by real-time RT-PCR. Furthermore, the effects of GDNF
treatment on cultured myenteric neurons (neuronal plasticity, receptor
expression) were monitored.
Results:
mRNA expression levels of GDNF and GFRa2 were down-regulated in the tunica
muscularis of patients with DD. LMD samples revealed high expression of GDNF
in circular and longitudinal muscle layers and high expression levels of the
corresponding receptors in myenteric ganglia. GDNF treatment of cultured
myenteric neurons increased mRNA expression of its corresponding receptors
and promoted neuronal plasticity revealed by synaptophysin mRNA expression.
Conclusions:
Our results suggest that the GDNF system is compromised in DD. In vitro studies
demonstrate that GDNF enhances mRNA expression of its receptors and
promotes enteric neuronal plasticity. Since patients with DD exhibit
hypoganglionosis and decreased expression of neuronal plasticity markers, we
propose that the observed neuronal loss in DD may be due to lacking
neurotrophic support mediated by GDNF.
Kategorie: Vortrag
Vortrag 9
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:3
Titel: Ankyrin-b in axons of the peripheral nervous system
Autoren: Engelhardt M.(1),Vorwald S.(1),Sobotzik J.(1),Gutzmann A.(1),Bennett,
V.(2),Schultz C.(1),
Adressen:(1)Med. Fakultät Mannheim der Universität Heidelberg|Neuroanatomie,
CBTM|Mannheim|Deutschland; email:maren.engelhardt@medma.uniheidelberg.de; (2)Duke University Medical Center|Howard Hughes Medical
Institute|Durhan, NC|USA
Abstract:
Ankyrin-B is a member of the ankyrin family of ubiquitously expressed membrane
adaptor proteins, which target diverse proteins to specialized axonal membrane
domains. In the central nervous system, ankyrin-B is specifically enriched in the
paranodal domain of myelinated axons. However, only little is known about the
distribution of ankyrin-B in the peripheral nervous system (PNS). Here we
document a conspicuous axonal distribution pattern of ankyrin-B identified in the
PNS of various mammalian species including rodents, pigs, non-human primates
and humans. The localization of ankyrin-B was analyzed using immunohistology
in combination with confocal laser scanning microscopy. Our analysis first
confirmed the previously reported localization of ankyrin-B in the paranodal
domain of thick myelinated axons localized in subcutaneous nerve bundles.
Surprisingly, ankyrin-B was also found in unmyelinated mechanosensory
terminals of type II (A beta) axons innervating Meissner or Pacinian corpuscles,
respectively. Anti-ankyrin-B labeling also permitted detailed visualization of the
mechanosensory terminals surrounding vibrissae. In the aforementioned
mechanosensory terminals, ankyrin-B exhibited a distinct membrane-associated
distribution. Finally, anti-ankyrin-B labeling provided an excellent and novel
method to visualize intraepidermal nerve fibers. These fibers are mainly
comprised of thin unmyelinated nociceptive C fibers. We thus hypothesize that
ankyrin-B may represent a constitutive component of unmyelinated PNS axons.
This was confirmed by the presence of ankyrin-B in unmyelinated fibers of the
sympathetic nervous system, innervating sweat glands, arrectores pilorum
muscles, and blood vessels. Taken together, these observations point to an
important role of ankyrin-B in somatosensory axon terminals and autonomic
nerve fibers of the PNS.
Kategorie: Vortrag
Vortrag 10
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:3
Titel: Age-dependent axonal trafficking of hcn1 channels in perforant path is
regulated by differential expression of trip8b isoforms
Autoren: Bender R.(1),Wilkars W.(1),Brandt N.(1),Chetkovich D.(2),
Adressen:(1)Universität Hamburg|Institut für Neuroanatomie|Hamburg|Germany;
email:rbender@uke.uni-hamburg.de; (2)Northwestern University Chicago|Dept.
Neurology & Clinical Neurosciences|Chicago|United States
Abstract:
The functions of HCN channels in neurons depend critically on their subcellular
localization, requiring fine-tuned machinery that regulates subcellular channel
trafficking. Here we provide evidence that regulatory mechanisms which govern
axonal HCN channel trafficking involve association of the channels with specific
isoforms of the auxiliary subunit TRIP8b. In the medial perforant path, which
normally contains HCN1 channels in axon terminals in immature but not in adult
rodents (Bender et al., J Neurosci, 2007), we found axonal HCN1 significantly
enhanced in adult mice fully deficient of TRIP8b (TRIP8b-/-). Interestingly, adult
mice only partially deficient of TRIP8b (Pex5l [1b-2] -/-), which still express major
TRIP8b isoforms 1a and 1a-4, did not show an altered HCN1 pattern compared to
wildtype, suggesting that presence of one or both of these isoforms prevents
HCN1 from being transported to the axons. In agreement with this hypothesis,
expression analyses demonstrated a strong increase of expression of both
TRIP8b isoforms in entorhinal cortex with age. However, when co-transfected
with HCN1 in immature entorhinal neuron culture, only TRIP8b (1a), but not
TRIP8b (1a-4), affected HCN1 subcellular distribution, resulting in
somatodendritic clustering of HCN1 channels and their reduced expression in
axons. Taken together, we conclude that TRIP8b isoforms are important
regulators of HCN1 trafficking in entorhinal neurons and that specifically isoform
TRIP8b (1a) could be involved in the age-dependent regulation of HCN1 channel
transport to perforant path axon terminals.
Kategorie: Vortrag
Vortrag 11
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:3
Titel: Regulation of chat expression by 17-beta-oestradiol in nsc-34 cells and in
the spinal cord
Autoren: Johann S.(1),Dahm M.(1),Beyer C.(1),
Adressen:(1)Universitätsklinikum, RWTH
Aachen|Neuroanatomie|Aachen|Deutschland; email:sjohann@ukaachen.de
Abstract:
Motoneurons in the spinal cord are the basic principle of cholinergic innervation of
the skeletal muscle. These neurons have also been shown to be exceedingly
affected in neurodegenerative disease like Amyotrophic lateral sclerosis (ALS).
The steroid hormone oestrogen (E) is known to act as a protective factor in the
central nervous system against neurodegenerative processes. Motoneuron
dysfunction often arises by the reason of alterations in the cholinergic system,
documented by a significant decrease in choline acetyltransferase (ChAT)
expression. Oestrogen has been shown to exert positive effects on cholinergic
neurons of the basal forebrain and the hippocampal formation. To expand that
work we have analyzed the effect of E on ChAT expression in the spinal cord. In
a first step we have established the presence of the E receptor alpha and beta in
NSC-34 cells, and in the cervical and lumbar parts of the mouse spinal cord.
Subsequently we investigated the effect of E treatment on ChAT expression in
vitro and in vivo. The application of E significantly increased the transcription of
ChAT mRNA in NSC-34 cells and in the cervical spinal cord. In addition we
confirmed the E effect on ChAT expression in the cervical spinal cord on protein
level. Our results indicate that E exhibits neurotrophic properties with respect to
the regulation of the cholinergic system by increasing ChAT expression in the
mouse spinal cord.
Kategorie: Vortrag
Vortrag 12
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:3
Titel: Gonadal steroid hormones protect from cortical damage and improve
functional recovery after ischemia
Autoren: Dang J.(1),Ulbrich C.(1),Kipp M.(1),Beyer C.(2),
Adressen:(1)RWTH Aachen Universität|Institut für
Neuroanatomie|Aachen|Deutschland; email:jdang@ukaachen.de; (2)RWTH
Aachen Universität|Institut für Neuroanatomie|Aache|Deutschland
Abstract:
Purpose: Ischemic conditions in the brain cause rapid and unrecoverable
neuronal damage accompanied by functional behavior dysfunction. Gonadal
steroids, estrogen (E) and progesterone (P), were found to ameliorate neuron
loss and cognitive disruption following traumatic brain injury. The present study
focuses on the short-term and long-term protective potential in a transient middle
cerebral artery occlusion model and aims to elucidate the underlying molecular
mechanisms.
Methods: Artery occlusion was performed with a filament for 1h and recovery was
tested after 23h and 14d. Hormones were applied through neck depots 1h after
stroke initiation, 12 and 24h thereafter. Hormone plasma values were monitored.
Young adolescent males and ovariectomized females were included in the study.
The infarct area was volumetrically measured, behavioral recovery tested using
standardized motor and cognitive tests, and gene/protein expression analyzed
from penumbra tissue.
Results: The application of E/P resulted in a reduction of the cortical infarct area
by approx. 60% in both sexes after day 1 and even after day 14 compared to
controls. Similar behavioral recovery rates were observed in both longitudinal
groups. Moreover, microglia and lymphocyte invasion in the penumbra are
massively attenuated and growth factors such as VEGF are up- and
proinflammatory markers such as the chemokines CCL2 and 5 and the cytokines
TNF-alpha and IL6 are down-regulated, respectively, after E/P application.
Conclusion: Our study shows that E/P application is neuroprotective in an
ischemic animal model. Further, we propose that the underlying neuroprotective
mechanisms include a mitigation of local inflammatory processes accompanied
by neovascularisation.
Kategorie: Vortrag
Vortrag 13
Rubrik: 2.Klinische Anatomie/Makroskopie
Abstract Nr.:2
Titel: Visualization of collagen fiber orientation by means of angle-dependent MRI
Autoren: Hammer N.(1),Seidel T.(1),Schneider G.(1),Garnov N.(2),Bechmann
I.(1),Steinke H.(1),
Adressen:(1)Universität Leipzig|Institut für Anatomie|Leipzig|Deutschland;
email:niels.hammer@medizin.uni-leipzig.de; (2)Universitätsklinikum Leipzig
AöR|Klinik und Poliklinik für Diagnostische und Interventionelle
Radiologie|Leipzig|Deutschland
Abstract:
Introduction: Many clinical problems related to ligaments and tendons are poorly
understood, because alterations in the respective tissues cannot be imaged well
with current MRI techniques. Here, we tested if angle-sensitive MRI offers
information on state and orientation of collagen fibers. Angle-sensitivity is based
on the magic-angle phenomenon in T2-weighted MRI scans between the collagen
fibers and the magnetic field (B0).
Material and Methods: T2-weighted MRI of three unfixed iliotibial tract specimens
were performed at different orientations: parallel (1), perpendicular (2), and
double oblique to B0 (3). The specimens were rotated stepwise from an angle
theta = 0° to 180° between the specimens and B0 with a step size of 10°. Voxelto-voxel correlation was performed between all images of the scan series, giving
the theta-intensity-curves for each voxel. An algorithm was developed for
calculating collagen fiber orientation from the theta-intensity-curves. The
correctness and robustness of the algorithm was validated using 1000
randomized virtual fibers. The calculated fiber orientation of the iliotibial tract
specimens was verified by polarization microscopy.
Results: Normalized angle-descriptive vectors were computed for each voxel.
With the virtual fibers, the mean deviation between actual and computed
orientation was 4°. Mean deviation of iliotibial tract orientation in polarization
microscopy and computation was 3° (1), 19° (2), and 10° (3).
Discussion: Angle-sensitive MRI is suitable to gain information on the main
orientation of collagen fibers within tissues.
Kategorie: Vortrag
Vortrag 14
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: Tubstain: a universal non-antibody tool to label microtubules
Autoren: Theiss C.(1),Neuhaus A.(2),Schliebs W.(2),Erdmann R.(2),
Adressen:(1)Ruhr-Universität Bochum|Institut für Anatomie und Molekulare
Embryologie|Bochum|Deutschland; email:carsten.theiss@rub.de; (2)RuhrUniversität Bochum|Abteilung für Systembiochemie, Institut für Physiologische
Chemie|Bochum|Deutschland
Abstract:
Imaging of the microtubular network is an important strategy to define cell-cycle
specific and pathological states of eukaryotic cells. Currently different
experimental approaches using fluorescence microscopy are used to visualize
and investigate the organization and dynamics of microtubules. Here we describe
TubStain, a novel recombinant tool allowing versatile and direct labelling of
microtubules in fixed cells over a broad variety of different species and tissues.
TubStain consists of three modules, a (i) Microtubule-binding domain based on
the sequence of human PEX14, a peroxisomal membrane protein which has
been recently identified as a membrane anchor for microtubules (Bharti et al,
2011, J. Cell Science), a (ii) dimerizer and a (iii) labelling agent. Conjugation with
different fluorophoretic proteins, dyes or gold-particles allows flexible, multiplexed
applications in various fields of microscopy, including fluorescence microscopy
and high-resolution electron microscopy. Direct microtubule visualization by
TubStain is faster and more reliable than convential immunostaining techniques
and could provide a suitable tool for identification and characterization of
microtubule polymerizing/depolymerizing drugs. Furthermore, the smallness of
TubStain in combination with the diversity of labelling agents makes TubStain an
ideal tool to study microtubules by high-resolution microscopic techniques.
Kategorie: Vortrag
Vortrag 15
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: Interplay of the cxcl12 / sdf-1 – binding chemokine receptors cxcr4 and
cxcr7 in human breast cancer cells
Autoren: Hattermann K.(1),Holzenburg E.(1),Hans F.(1),Held-Feindt
J.(2),Mentlein R.(1),
Adressen:(1)Christian-Albrechts-Universität zu Kiel|Anatomisches
Institut|Kiel|Deutschland; email:k.hattermann@anat.uni-kiel.de; (2)ChristianAlbrechts-Universität zu Kiel|Klinik für Neurochirurgie UKSH Campus
Kiel|Kiel|Deutschland
Abstract:
The chemokine CXCL12 / stromal cell-derived factor-1 (SDF-1) is a small soluble
peptide that is known to promote breast cancer metastasis via its long renown
receptor CXCR4. Recently, a novel seven-transmembrane receptor for CXCL12
was discovered and named CXCR7. Initially, this receptor was supposed to be a
decoy receptor as it lacks the conserved DRY motif that is necessary for Gprotein coupling. However, interested in the interplay of both receptors we
investigated their transcription in four different mamma carcinoma cell lines by
qRT-PCR followed by native fluorescence-immunocytochemistry. One selected
cell line (MCF-7) expressed both receptors at comparable level. They were
localized on the cell surface and were internalized upon stimulation with CXCL12.
Looking for functional effects that were directly mediated via CXCR7 we analyzed
signal transduction (Western blot on phosphorylated kinases) and anti-apoptotic
effects (e.g. Nicoletti-staining) using selective antagonists for CXCR4 or CXCR7.
We found out that CXCL12 can signal and mediate functional, in particular antiapoptotic effects directly via CXCR7. Thus, CXCR7 is a functional receptor in
human mamma carcinoma cells and is not merely acting as a regulator of CXCR4
signaling by scavenging extracellular CXCL12.
Kategorie: Vortrag
Vortrag 16
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: The new lymphangiogenesis inhibitor esvegfr2 is down-regulated in
advanced stage neuroblastoma and induced upon differentiation.
Autoren: Becker J.(1),Wilting J.(2),
Adressen:(1)Universitätsmedizin Göttingen|Zentrum Anatomie, Abt. Anatomie
und Zellbiologie|Göttingen|Deutschland; email:juergen.becker@med.unigoettingen.de; (2)Universitätsmedizin Göttingen|Zentrum Anatomie Abt. Anatomie
und Zellbiologie|Göttingen|Deutschland
Abstract:
Tumor metastasis is facilitated by hemangiogensis and lymphangiogenesis. Key
regulators of angiogenesis are the members of the vascular endothelial growth
factor (VEGF) family and their receptors (VEGFR). Recently, an endogenous
soluble VEGFR-2 splice variant (esVEGFR-2) was described (Albuquerque et al.
2009). It does not bind VEGF-A, the hemangiogenic ligand of the membranebound variant, but rather VEGF-C, the key inducer of lymphangiogenesis.
We investigated esVEGFR2 in neuroblastoma (NB), the most common solid
extracranial malignancy of childhood. NB is an embryonic tumour, derived from
neural crest descending sympatho-adrenal progenitor cells. In clinical NB staging,
infestation of distant lymph nodes is a critical sign, demanding classification into
the most advanced stage 4.
We present that expression of esVEGFR-2 is lower in the progressed stages 3
and 4. We also found that MYCN amplification, the most adverse prognostic
marker in NB, correlates with lower esVEGFR-2 expression. In human embryonic
tissue we demonstrate expression in sympathetic ganglia and the adrenal
medulla, indicating that esVEGFR-2 contributes to normal development of
sympatho-adrenal organs. Examination of esVEGFR-2 in NB cells after treatment
with the differentiating agent all-trans retinoic acid (ATRA) reveals that ATRAinduced differentiation enhances esVEGFR-2 expression.
Tumor angiogenesis is not only achieved by the up-regulation of pro-angiogenic
molecules (VEGF-A, VEGF-C) but also by the down-regulation of inhibitory
molecules like esVEGFR-2. Additionally, esVEGFR-2 is associated with normal
development of sympatho-adrenal tissues and can be induced in NB cell lines by
differentiating ATRA treatment. Therefore sVEGFR-2 may be a potent regulator
of lymphangiogensis in both normal development and tumors.
Kategorie: Vortrag
Vortrag 17
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: Nuclear stem cell factor hmga2 utilizes chk1 signaling to suppress apoptosis
in human cancer cells
Autoren: Klonisch T.(1),Natarajan S.(2),Hombach-Klonisch S.(2),Ghavami
S.(3),Droege P.(4),
Adressen:(1)University of Manitoba|Dept. of Human Anatomy and Cell
Science|Winnipeg|Canada; email:klonisch@cc.umanitoba.ca;
(2)UoManitoba|Human Anatomy and Cell Science|Winnipeg|Canada;
(3)UoManitoba|Dept. of Physiology|Winnipeg|Canada; (4)Nanyang Technological
University|School of Biological Sciences|Singapore|Singapore
Abstract:
Introduction: The non-histone chromosomal protein High Mobility Group AT-hook
protein-2 (HMGA2) is expressed in embryonic stem/progenitor cells, not present
in normal adult cells but re-expressed in cancer cells. HMGA2 is a marker of the
aggressive
undifferentiated
thyroid
cancer
(UTC)1
and
increases
chemoresistance of tumor cells by promoting Base Excision Repair (BER)2. Upon
DNA damage, DNA damage repair signaling factors like the nucleoplasmic
checkpoint kinases CHK1 and CHK2 are activated/ phosphorylated by ATR and
ATM kinase, respectively.
Hypothesis: HMGA2 protects cells from DNA damage-induced cell death.
Results: Upon MMS-induced DNA damage, HMGA2 clones displayed
significantly less nuclear immunoreactive gamma-H2AX and RPA34 and shorter
tail moments in comet assays as compared to mock clones. MMS insult resulted
in a increased and sustained presence of pCHK1S296 even 24 h after MMS
induced DNA damage in HMGA2 clones, but not mock transfectants. This
coincided with G2/M arrest in HMGA2 clones after MMS insult. Similar results
were obtained with HT1080 fibrosarcoma cells in which endogenously produced
HMGA2 could be suppressed by a doxycycline-inducible shHMGA2 construct.
Thus, sustained phosphorylation of CHK1 at residue serin296 required the
presence of HMGA2. MMS treatment of UTC with specific knock-down of CHK1
caused an increase in apoptosis in HMGA2 transfectants. Similar results were
obtained for HMGA2 expressing A549 lung cancer transfectants.
Conclusion: We identified HMGA2 to mediate the activation of the CHK1 signaling
cascade, cause prolonged G2/M arrest, and enhance Base Excision Repair.
Thus, HMGA2 caused increased DNA damage repair and inhibited DNA damageinduced apoptosis in HMGA2+ human cancer cells.
Kategorie: Vortrag
Vortrag 18
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: Chemotherapy of brain cancer: a novel approach using rhcc microcarriers
Autoren: Thanasupawat T.(1),Hombach-Klonisch S.(2),Krcek J.(3),Del Bigio
M.(4),Stetefeld J.(5),Klonisch T.(2),
Adressen:(1)UoManitoba|Department of Human Anatomy and Cell
Science|Winnipeg|Canada; (2)UoManitoba|Human Anatomy and Cell
Science|Winnipeg|Canada; (3)UoManitoba|Surgery|Winnipeg|Canada;
(4)UoManitoba|Pathology|Winnipeg|Canada;
(5)UoManitoba|Chemistry|Winnipeg|Canada; email:klonisch@cc.umanitoba.ca
Abstract:
Introduction: Treatment for Glioblastoma (GB), a common and most aggressive
form of primary brain cancer, includes surgery, radiation, and chemotherapy.
Chemoresistance and severe irreversible toxic side effects are complications of
cisplatin (CP) treatment. Gold compounds are new anticancer drugs. We
developed a novel microcarrier system for the delivery of CP and gold
compounds into cancer cells and treatment of brain cancer. Right Handed Coiled
Coil (RHCC) is a highly stable helical protein and originates from the
archaebacterium Staphylothermus marinus. The crystal structure of RHCC
revealed four large hydrophobic cavities inside tetrameric tubes capable of storing
CP and gold compounds. We created a Carrier-Pathfinder-System by linking
RHCC to an EGF-affibody for EGFR-targeted receptor binding on GB cells.
Further, we coated RHCC with polyethylene glycol (PEG) compounds to assist in
the delivery RHCC into GB cells.
Hypothesis: RHCC are effective delivery systems for the treatment of brain
cancer.
Results: GB cells readily take up Alexaflour488nm–labelled RHCC (AF-RHCC)
and PEG coating enhanced the cellular uptake of RHCC. Low concentrations of
RHCC laden with CP and gold (CP-/Au-RHCC) microcarriers were vastly superior
in killing primary GB cells than free CP/Au compounds.
Conclusion: Our unique coiled-coil microcarrier system provides a novel strategy
for efficient drug delivery of chemotherapeutic agents into tumour cells, reducing
the toxic side effects and resulting in superior therapeutic potential in the
treatment of brain cancer patients.
Kategorie: Vortrag
Vortrag 19
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: Nrf2 is crucial for liver regeneration
Autoren: Wruck C.(1),Streetz K.(2),Fragoulis A.(1),Rosen C.(1),Pufe T.(1),
Adressen:(1)RWTH Aachen|Institut für Anatomie und
Zellbiologie|Aachen|Deutschland; email:cwruck@ukaachen.de; (2)RWTH
Aachen|Klinik für Gastroenterologie|Aachen|Deutschland
Abstract:
Background: Aim of this study was to elucidate role of the redox-sensitive
transcription factor Nrf2 in hepatic regeneration.
Methods: Nrf2-knockout and hepatic Keap1-knockout mice, which have a
hepatocyte specific overactive Nrf2, were feed on 3,5-diethoxycarbonyl-1,4dihydrocollidine (DDC) containing diet resulting in chronic cholestatic liver injury
or were hepatectomised to study regeneration processes.
Results: Mice deficient in Nrf2 showed significant more lymphocyte infiltration and
cytokine expression as wild type mice. Significantly more necrosis, apoptosis and
cholestasis became evident in Nrf2-knockout mice. This was associated with
stronger periportal oval cell activation. In contrast, mice with hepatocyte specific
knockout of Keap1, the inhibitor of Nrf2, showed significant less oxidative liver
damage, and less lymphocyte infiltration. Interestingly, Keap1-liver knockout
leads to enhanced stem cell proliferation in response to DDC feeding. Moreover,
we show that Nrf2 is crucial for the up-regulation of Multidrug Resistance-Related
Proteine 2 and 3 that protects hepatocytes against various toxins.
Conclusion: We show that Nrf2-dependent signalling pathways are important to
protect the liver against chronic cholestatic liver injury. In addition, Nrf2 is
important for the activation of oval cells that accomplish liver regeneration. These
findings indicate that the use of Nrf2-inducers might be considered as a novel
therapeutic strategy to combat hepatic diseases and accelerate liver
regeneration.
Kategorie: Vortrag
Vortrag 20
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: Regulation of mediators associated with osteoarthritis by tnfα in human
synovial fibroblasts and in co-culture with chondrocytes in vitro
Autoren: Schulze-Tanzil G.(1),Mrosewski I.(1),Conrad C.(1),Jork N.(1),Wiegand
E.(1),Gorthe K.(1),Ertel W.(1),John T.(1),
Adressen:(1)Charité Universitätsmedizin, CBF|Klinik für Unfall- und
Wiederherstellungschirurgie|Berlin|Deutschland; email:gundula.schulzetanzil@charite.de
Abstract:
Osteoarthritis (OA) is usually associated with synovitis. Pro-inflammatory
mediators such as tumor necrosis factor (TNF)alpha are secreted by several cell
types within the osteoarthritic joint and might affect synovial fibroblasts (SF).
Hence, we analyzed the impact of TNFalpha on osteoarthritic SF and
characterized the interplay between articular chondrocytes and SF in an
interactive direct co-culture model.
Primary human SF were isolated from the knee joint synovium and tested for their
typical protein expression profile (CD55+, CD44+, UDPGDH+, Tenascin+, CD14). For gene expression analysis of OA associated mediators such as TNFalpha,
IL-6, MMP1, and -3, SF were stimulated with TNFalpha, IL-10 or TNFalpha + IL10 (each 10 ng/mL) for 24 h. A three-dimensional dynamical co-culture model
was established using porcine chondrocytes pellets and a human synovial cell
line to analyse the effect of TNFalpha on both cell types on gene expression and
histology.
Cultured SF were activated by TNFalpha alone or in combination with IL-10 and
up-regulated their mRNA-expression for typical immunoregulatory and catabolic
mediators. Treatment of chondrocytes pellets with TNFalpha, co-culturing of
chondrocytes with SF with or without TNFalpha stimulation lead to cartilage
degradation and suppression of type II collagen and aggrecan gene expression.
These findings might underline the active role of SF in the pathogenesis of OA
and suggest them as a potential therapeutic target. An ongoing study, using
overexpression vectors for human and viral IL-10 in SF, could show whether the
SF activation by TNFalpha can be modulated by high local levels of IL-10.
Kategorie: Vortrag
Vortrag 21
Rubrik: 5.Entwicklungsbiologie
Abstract Nr.:5
Titel: Dynamics of molecular factors during mammalian germ cell specification
Autoren: Püschel B.(1),Hopf C.(1),Muske M.(1),Siede J.(1),Bosch A.(1),Viebahn
C.(1),
Adressen:(1)Georg-August-Universität Göttingen|Zentrum Anatomie, Abteilung
Anatomie und Embryologie|Göttingen|Deutschland; email:bpuesch@gwdg.de
Abstract:
Germ cell founders segregate early during animal development from the somatic
tissues of the embryo. In contrast to the preformation mechanism found in many
species, the germ cell forerunners in mouse and probably all other mammals are
selected during gastrulation by an induction process intricately coordinated in
time and space and involving, for example, the transcription repressors PRDM1,
PRDM14, the pluripotency associated homeobox transcription factor NANOG and
extracellular signals. To find general mechanisms of mammalian germline
specification we analyzed the spatial and temporal mRNA expression patterns of
the transcriptional regulators. In situ hybridized rabbit embryos were examined
using light microscopy and were compared to immune fluorescence stained
embryos labeled with the germ cell specific antibody directed against the
mitochondria-associated antigen PG2. The expression patterns of PRDM1 and
NANOG are in agreement with the respective position of PG2-stained primordial
germ cells (PGCs) and correspond to the expression patterns observed in
homologous stages of the mouse. In contrast to mouse, however, PRDM14 was
not found in rabbit PGCs, at least until the early somite stages. Quantification of
the stained PGCs revealed a high variability when comparing different embryos at
any given developmental stage but there is a steady increase when PGC
numbers are plotted against developmental time. The differences in the total
PGCs counts per embryo with regard to the three markers examined here might
be due to dynamic mRNA expression changes needed for the molecular control
of germ cell specification and maintenance.
Kategorie: Vortrag
Vortrag 22
Rubrik: 5.Entwicklungsbiologie
Abstract Nr.:5
Titel: Interruption of caudorostral diffusion in the presomitic mesoderm affects
somite formation in the avian embryo
Autoren: Huang R.(1),Pu Q.(1),Regen W.(2),Christ B.(2),
Adressen:(1)University of Bonn|Institute of Anatomy|Bonn|Germany;
email:ruijin.huang@uni-bonn.de; (2)University of Freiburg|Institute of Anatomy
and Cell Biology|Freiburg|Germany
Abstract:
The metameric pattern of the axial skeleton is established by somite formation
from the presomitic mesoderm(PSM), so-called somitogenesis. The positioning of
the somite boundaries is specified by the wavefront which is identified as a
traveling threshold of FGF signaling gradient in the PSM. Disrupting the FGF
signaling gradient by grafting beads soaked in FGF8 results in an anterior shift of
the wavefront leading to the formation of smaller somites. In contrast, blocking
FGF signaling displaces the front caudally and causes the formation of bigger
somites. The FGF signaling gradient is believed to be generated by a gradient of
Fgf8 mRNA, which is translated into a gradient of signaling protein across the
PSM. It is still unknown to what extent the diffusion of FGF8 protein contributes to
the formation of FGF8 gradient in the PSM. In this study, we address the
importance of the diffusion mechanism during somitogenesis by blocking the
caudorostral protein diffusion in the PSM. After implanting an impermeable
barrier, the graded fashion of FGF signaling detected by the phosphorylated form
of MAPK is altered. As a result, somites located rostrally to the barrier are bigger
than their equivalents on the control side. The vertebra half of the manipulated
side is accordingly bigger than the half of the control side. This effect can be
repealed by implantation of a FGF8-bead anterior to the barrier. These results
suggest that diffusion is involved in the establishment of the FGF signaling
gradient in the PSM during somitogenesis.
Kategorie: Vortrag
Vortrag 23
Rubrik: 5.Entwicklungsbiologie
Abstract Nr.:5
Titel: Nestin reveals progenitor cells in pulmonary vasculature.
Autoren: Saboor F.(1),Berndt C.(1),Weissmann N.(2),Müller D.(1),
Middendorff R.(1),
Adressen:(1)Justus Liebig University, Gießen|Institute of Anatomy and Cell
Biology|Gießen|Germany; email:farhan.saboor@anatomie.med.uni-giessen.de;
(2)Justus Liebig University, Gießen|Excellence Cluster Cardio Pulmonary
Systems (ECCPS)|Gießen|Germany
Abstract:
Vascular smooth muscle cells (VSMCs) and pericytes (PCs), distinguished by the
expression of neuronal stem cell marker “Nestin”, may represent stemcell-like
progenitor cells for tissues in various organs. In one of our previous studies, we
found that nestin-expressing VSMCs and PCs in testicular blood vessels are the
progenitors of testosterone producing Leydig cells.
To analyze the expression pattern of nestin and its role as marker for proliferating
progenitor cells in the lung, nestin expression and localization was investigated
during postnatal development in nestin-GFP mice. To investigate nestin
expression during vascular remodelling, samples from two models of pulmonary
hypertension (PH) [monocrotaline (MCT) rat model and hypoxic mouse model] as
well as human samples from patients of PH were analyzed. Nestin data was
compared with expression of proliferation markers (PCNA, Ki67) and PDGF
receptors.
Nestin was found in a subpopulation of VSMCs in lung vasculature. As compared
to adult normoxic controls significantly higher nestin expression was observed in
pulmonary vasculature of postnatal tissues and in adult lungs between day 3-7 of
hypoxic exposure but not at later time points when PH became evident. Increase
of nestin correlated well with an increase of cell proliferation. In hypoxic lungs
peak of phosphorylated (activated) PDGF receptor-ß correlated with nestin one.
Increase of nestin-immunoreactive VSMCs was also found in MCT rat and human
lung samples.
Certain contractile cells capable of proliferation could be identified by nestin
expression in lungs and may be used as prognostic marker and new target for
therapeutic interventions of diseases like PH.
Kategorie: Vortrag
Vortrag 24
Rubrik: 5.Entwicklungsbiologie
Abstract Nr.:5
Titel: Expression and function of the sdf-1 chemokine receptors cxcr4 and cxcr7
during limb muscle development and regeneration
Autoren: Ödemis V.(1),Hunger C.(1),Engele J.(1),
Adressen:(1)Leipzig|Anatomie|Leipzig|Deutschland;
email:Veysel.Oedemis@medizin.uni-leipzig.de
Abstract:
The chemokine stromal cell derived factor-1 (SDF-1/CXCL12), and its receptor,
CXCR4, control distinct steps of limb myogenesis. Recently, CXCR7 was
identified as a second receptor for SDF-1. Since little is known about the role of
CXCR7 in skeletal muscle development, we have now investigated the
expression of CXCR7 in comparison to CXCR4 in developing and regenerating
limb muscles. Additionally, we have analyzed whether or not CXCR7 actively
mediates SDF-1-signalling and influences myogenic differentiation of C2C12
myoblasts. Treatment of C2C12 cells, which co-express CXCR4 and CXCR7,
with SDF-1 activated Erk and PKCzeta and inhibited myogenic differentiation.
Inactivation of CXCR4 and CXCR7 by antagonists and siRNA showed that only
CXCR4-depletion abrogated SDF-1-dependent cell signalling and subsequent
inhibitory influences on myogenic differentiation. We further found that in vivo
SDF-1 is continuously expressed by the endomysium of postnatal and adult limb
muscles. Interestingly, CXCR4 expression is highest in late embryonic limb
muscles and drops shortly after birth when secondary muscle growth terminates.
By contrast, CXCR7 expression increases perinatally in limb muscles and
persists into adult life. Analysis of regenerating muscles of adult dystrophindeficient mdx mice showed re-expression of CXCR4, whereas expression of
CXCR7 remained unchanged. Collectively, these findings point to a crucial role of
SDF-1 and its receptors, CXCR4 and CXCR7, in muscle development and
regeneration.
Kategorie: Vortrag
Vortrag 25
Rubrik: 5.Entwicklungsbiologie
Abstract Nr.:5
Titel: Atoh8, a novel regulator of skeletal myogenesis and muscle regeneration
Autoren: Balakrishnan-Renuka A.(1),Balakrishnan-Renuka A.(1),Yusuf F.(1),Patel
K.(2),Otto A.(2),Morosan-Puopolo G.(1),Chen J.(1),Theiss C.(1),Chankiewitz
V.(1),Zoidl G.(1),Philippi S.(3),Dai F.(3),Brand-Saberi B.(1),
Adressen:(1)Ruhr University Bochum|Institute of Anatomy, Department of
Anatomy and Molecular Embryology|Bochum|Germany; (2)University of
Reading|School of Biological Sciences|Reading|Uk; (3)University of
Freiburg|Institute of Anatomy and Cell Biology|Freiburg|Germany;
email:beate.brand-saberi@rub.de
Abstract:
Skeletal myogenesis and myogenic regeneration are essentially very similar
processes that ensure that proper functional muscle tissue is formed during
development and maintained in the course of postnatal life. Satellite cells being
the main source of resident muscle stem cells are mainly responsible for the
extensive muscle growth during late embryonic development and also for muscle
regeneration in adult life. We show that ATOH8 is expressed in the somite of
chicken embryos and silencing of ATOH8 in chicken somites perturbs skeletal
myogenesis. Furthermore, we show here for the first time that ATOH8, a bHLH
transcription factor is expressed along with Pax7 in satellite cell as well as in
skeletal myoblasts. Our results show that ATOH8 is expressed in activated
satellite cells and is downregulated as cells enter terminal differentiation.
Regenerating muscle shows an upregulated ATOH8 expression at site of injury.
Preliminary data from our protein studies show, for the first time, that cytoplasmic
ATOH8 interacts with the catalytic subunit of Calcineurin (CnA beta). Our results
are the earliest report showing the involvement of ATOH8 in embryonic
myogenesis and satellite cell differentiation. We conclude that ATOH8 is essential
for the fine regulation of the essential balance between skeletal myogenesis and
self renewal of satellite cell.
Kategorie: Vortrag
Vortrag 26
Rubrik: 5.Entwicklungsbiologie
Abstract Nr.:5
Titel: Spatial synexpression of dynein and nodal in the early notochord of the
mammalian embryo: breaking left-right -symmetry without ciliary flow?
Autoren: Schröder S.(1),Weizbauer A.(1),Tsikolia N.(1),Beckers A.(2),Gossler
A.(2),Viebahn C.(1),
Adressen:(1)Georg-August-Universität|Zentrum Anatomie, Abteilung Anatomie
und Embryologie|Göttingen|Deutschland; email:silke.s.schroeder@hotmail.de;
(2)Medizinische Hochschule Hannover|Institut für
Molekularbiologie|Hannover|Deutschland
Abstract:
Left-sided nodal expression in the lateral plate mesoderm (LPM) fixes the
molecular left-right (LR) differentiation of the mammalian embryo at the 4-somite
stage. The preceding labile phase of symmetry breaking is marked by
paramedian nodal expression in lateral cells of the posterior notochord (PNC),
which is either restricted to the left from the start (pig, cow), or is symmetrical at
first and then thought to be lateralized to the left through ciliary flow on the free
ventral surface of the PNC (mouse, rabbit). However, recent findings obtained in
mice mutant for pkd1I1 and pkd2 genes, coding for a mechanosensory receptor
complex, revealed severe LR defects in the presence of normal flow. Apart from a
theoretical support for the \"two-cilia hypothesis\", these results raise the question
as to a flow-independent, intracellular symmetry breaking mechanism, which may
involve dynein axonemal heavy chains (DNAHs), ciliary motorproteins associated
with LR defects. Our high-resolution in situ hybridisation analysis show the
expression of several DNAHs in mouse and rabbit to be confined to the node, and
later to the PNC. At stage 6 (late pre-somite) and 7 (1-3 somite pairs) DNAH
expression correlates with the domain of rotating PNC cilia and lies at the level of
the paramedian (symmetrical) nodal expression in lateral PNC cells, thus
separating this domain directly. This spatial synexpression of DNAHs and nodal
within the PNC lends support to a hypothesis which assumes a cilia-independent,
intracellular planar symmetry breaking function of DNAHs and may be universal
for amniotes.
Kategorie: Vortrag
Vortrag 27
Rubrik: 5.Entwicklungsbiologie
Abstract Nr.:5
Titel: The node breaks l-r symmetry through notochord formation in the chick
embryo.
Autoren: Tsikolia N.(1),Schwartz P.(1),Viebahn C.(1),
Adressen:(1)Göttingen|Zentrum Anatomie, Anatomie und
Embryologie|Göttingen|Deutschland; email:nikoloz.tsikolia@med.unigoettingen.de
Abstract:
A recent timeline analysis of symmetry breaking in chick embryo made nodal the
first asymmetrical molecular marker, expressed at the early notochord stage
already. However, it remains to be determined whether morphological asymmetry
in the node occurs beforehand and whether paranodal leftward cell movements
lead to, or follow this morphological asymmetry. This study defined staging
criteria concerning node and notochord formation in chick embryos by correlating
high-resolution light microscopy and scanning electron microscopy. The node at
stage 4 shows a primitive pit and symmetrical lateral margins, here termed
“shoulders”. Signs of epithelial-mesenchymal transition in semithin sections are
first detectable in the node at stage 4+, which is defined by increased density of
the mesoderm anterior to the node; at the same stage, the right shoulder begins
to be thicker than the left one. At stage 5-, defined by the appearance of epithelial
(notochord) differentiation in the mesoderm compartment anterior to the node,
epiblast cells in the left shoulder are columnar while right shoulder cells appear
small and mesenchymal; scanning electron microscopy and spot-light illumination
show that the right shoulder is tilted to the left (“leftward torsion”). DiI labeling of
right paranodal tissue revealed cellular displacement mainly towards the anterior
but not crossing the midline. Thus, LR-differences in the node start at stage 4, i.e.
prior to asymmetrical nodal expression, and suggest that the initiation site of
notochord formation is shifted to the right. The results underline the tight temporal
connection between node asymmetry and notochord formation.
Kategorie: Vortrag
Vortrag 28
Rubrik: 5.Entwicklungsbiologie
Abstract Nr.:5
Titel: A cell-autonomous role for ciliary IFT proteins in axonal guidance in the
embryonic spinal cord
Autoren: Tucker K.(1),Wilson N.(2),Stoeckli E.(2),
Adressen:(1)Universität Heidelberg|Anatomie und medizinische
Zellbiologie|Heidelberg|Deutschland; email:kerry.tucker@urz.uni-hd.de;
(2)Universität Zürich|Institute of Molecular Life Sciences|Zürich|Schweiz
Abstract:
Primary cilia are crucial for transduction of Sonic hedgehog (Shh) signaling in
many organs throughout the body during embryogenesis, including the central
nervous system. Antero- and retrograde transport within cilia is accomplished by
a small family of intraflagellar transport (IFT) proteins, whose role in Shh signaling
has been thought to be restricted to the primary cilium itself. We have uncovered
a novel role of IFT88 and IFT52 in the guidance of commissural axons of the
spinal cord in both mouse and chick embryos. In the mouse at embryonic day (E)
12.5, a hypomorphic mutant of IFT88, cobblestone, displayed both stalling and
random growth of commissural axons at the midline. This correlated with a loss of
Shh expression and a disrupted floorplate morphology. In the chick, bi- or
unilateral knockdown of IFT88 by in ovo RNAi at E2 or E3 resulted in similar
phenotypes, with axons also failing to make a turning decision at the contralateral
floor plate border. Similar results were obtained upon RNAi directed against
IFT52. Notably, targeted electroporation of IFT88 dsRNA into the ventral spinal
cord did not affect pathfinding decisions of the dorsally-located commissural
neurons. In contrast to the mouse, spinal cord patterning markers were not
affected following unilateral knockdown of IFT88 at E3. Together, these results
suggest that normal IFT protein function is necessary within the commissural
neurons themselves for proper Shh-directed outgrowth. Whether the activity of
IFT proteins is restricted to the primary cilia of these neurons is under
investigation and will be discussed.
Kategorie: Vortrag
Vortrag 29
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: The effect of mycotoxin deoxynivalenol and e.coli lipopolysaccharide on
small intestinal morphology and zonula occludens-1 protein in vivo.
Autoren: Klüß J.(1),Klunker L.(1),Walk N.(1),Nossol C.(1),Kahlert S.(1),Brosig
B.(2),Döll S.(2),Dänicke S.(2),Rothkötter H.(1),
Adressen:(1)Otto-von-Guericke Universität|Institut für
Anatomie|Magdeburg|Deutschland; email:jeannette.kluess@med.ovgu.de;
(2)Bundesforschungsinstitut für Tiergesundheit|Institut für
Tierernährung|Braunschweig|Deutschland
Abstract:
The mycotoxin deoxynivalenol (DON) and E.coli lipopolysaccharide combined are
suspected to have a synergistic effect on intestinal architecture and epithelial
barrier integrity. We investigated the proposed effect of DON and LPS on crypt
depth and zonula occludens-1 protein (ZO-1) in the porcine small intestine.
The study comprised six experimental groups (6 x 6 pigs). Animals were fed a
control or a diet containing DON for four weeks. Subsequently, control group was
infused for an hour either with 100µg/kg BW DON, 7.5µg/kg BW LPS, a
combination of both or 0.9% NaCl and the DON group with LPS or NaCl only.
Animals were sacrificed 3.25 hours after start of infusion. Tissue samples were
taken from duodenum, proximal jejunum, mid-jejunum, proximal ileum and
terminal ileum. Crypt depth was measured and expression of ZO-1 analysed by a
scoring system. Data were analysed by repeated-measures using ANOVA and
Dunnett’s post hoc test.
A physiological decrease of crypt depth along the proximo-distal axis was
observed but no difference between control and any experimental groups. The
structural organisation of ZO-1 was severely damaged in the mid jejunum in
animals fed a control diet and infused with DON only. There was no difference to
control in any other intestinal region or experimental group. LPS did not affect any
of the investigated parameters, thus a synergistic effect of DON and LPS was not
present.
Our data are consistent with previous work showing the damaging effect of DON
on ZO-1 structure in an in vitro cell culture system with polarised intestinal porcine
epithelial cells.
Kategorie: Vortrag
Vortrag 30
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: Staphylococcus aureus septic isolates evolve different pathogenic strategies
to target endothelial barrier function
Autoren: Kramko N.(1),Seebach J.(1),Löffler B.(2),Heilmann C.(2),Peters
G.(2),Schnittler H.(1),
Adressen:(1)Westfälische-Wilhelms Universität|Institut für Anatomie und
Vaskuläre Biologie|Münster|Deutschland; email:kramko@uni-muenster.de;
(2)Westfälische-Wilhelms Universität|Institut für Medizinische
Mikrobiologie|Münster|Deutschland
Abstract:
The Gram-positive bacterium Staphylococcus aureus is a life-threatening
pathogen that causes systemic endovascular infections frequently accompanied
with the development of sepsis and septic shock syndrome. We aimed to unravel
early barrier function-related pathogen spread, a critical pathophysiological step
in these severe diseases. Using primary human endothelial cells that were
infected with eight different septic isolates of S. aureus, we investigated the two
potential spreading pathways, the paracellular spreading and transcellular
spreading. Thus, we combined measurements of the transendothelial electrical
resistance, a parameter that gives information of paracellular barrier function,
activation and transwell-filter assay that allows the determination of translocated
bacteria through the endothelial cell layer. Six of the eight strains significantly
decreased the paraendothelial barrier function by more than 20% within 3 hours.
This was associated with certain degrees of activation as tested by FACS
analyses after ICAM1 surface staining. In contrast, two strains had nearly no
effect on TER, and one of them showed a high translocation rate but did not show
any activation. The data show for the first time both strain-dependent breakdown
in paraendothelial barrier function and bacterial translocation across the
endothelial layer. We propose that bacterial translocation might be blocked by
endothelial activation.
Kategorie: Vortrag
Vortrag 31
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: Epithelial protein lost in neoplasm, eplin, is expressed in endothelial cells
and colocalizes with actin filaments at lammellipodia and at cell junctions
Autoren: Taha M.(1),Abu Taha A.(1),Schnittler H.(1),
Adressen:(1)Westfälische Wilhelms-Universität Münster|Dept. of Anatomy and
Vascular Biology|Münster|Deutschland; email:m_taha01@uni-muenster.de
Abstract:
Adherens junctions of endothelial cells consist of the VE-cadherin/catenin
complex that is directly or indirectly connected to junction-associated actin
filaments. The molecular interaction of the VE-cadherin-catenin complex and the
actin filamnts is not completely understood and is controversially discussed.
Recently, the Epithelial Protein Lost In Neoplasm¡ EPLIN, has been indicated to
be the missing link between the E-cadherin-catenin complex and junction
associated actin filaments in epithelial cells and might further control the actin
dynamics by interaction with the Arp2/3 complex. Here we show by Western blot
analyses and immunofluorescence microscopy that EPLIN is expressed in
cultured human umbilical vein endothelial cells and in endothelium in vivo.
Immuno-fluorescence staining localizes EPLIN at lamelliopodia of cultured
endothelial cells in colocalization with the Arp 2/3 complex and with actin
filaments. In addition EPLIN was localized at junctions of confluent endothelial
cells in colocalization with VE-cadherin, catenins and junction associated actin
filaments. In addition, alpha and beta- EPLIN-isoforms were tagged to MCherry
and EGFP respectively and expressed in endothelial cells by lentiviral gene
transfer. By live cell imaging using high speed spinning disc microscopy we
demonstrate high dynamic activity of EPLIN-isoforms at lammellidodia and at the
cell junctions. The data indicate that EPLIN is a critical component of
lammellipodia and cell junctions in endothelial cells.
Kategorie: Vortrag
Vortrag 32
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: Akap-mediated pka compartmentalization contributes to formation and
functional maintenance of the endothelial barrier
Autoren: Radeva M.(1),Spindler V.(1),Waschke J.(1),
Adressen:(1)Ludwig-Maximilians-University|Institute of Anatomy and Cell Biology.
Department 1|Muenchen|Germany; email:Jens.Waschke@med.uni-muenchen.de
Abstract:
Aim: Maintenance of the endothelial barrier is mediated by cAMP-dependent
signaling pathways which at least in part involves protein kinase A (PKA). Tight
regulation of PKA function can be achieved by discrete compartmentalization of
the enzyme via physical interaction with A-kinase anchoring proteins (AKAPs). By
using a cell-permeable synthetic peptide (TAT-Ahx-AKAPis) designed to
competitively inhibit PKA-AKAP interaction, we investigated the importance of
AKAP function for cAMP-dependent maintenance of the endothelial barrier in vivo
and in vitro. Methods and Results: Analysis of human (HDMEC) and mouse
(MyEnd) microvascular endothelial cells as well as isolated rat mesenteric
microvessels was performed. In vivo microvessel hydraulic conductivity (Lp)- and
in vitro transendothelial electrical resistance (TER) measurements showed that
TAT-Ahx-AKAPis destabilized endothelial barrier properties. Moreover, due to
TAT-Ahx-AKAPis addition, barrier stabilization in response to increased cAMP
following treatment with forskolin/rolipram (F/R) was also blunted in vitro.
Immunofluorescence analysis revealed that TAT-Ahx-AKAPis induced
reorganization of both, adherens junctions and the actin cytoskeleton. Those
effects were paralleled by decrease in the intensity of PKA and Rac1 membrane
staining as well as by Rac1 inactivation. Similarly, membrane localization of
AKAP220, which was found to interact with the endothelial cadherin-catenin
complex, was also reduced. In addition, as evaluated by TER measurement,
AKAP220 siRNA knockdown significantly impaired endothelial barrier formation.
Conclusions: Taken together, these results indicate that AKAP-mediated PKA
subcellular compartmentalization and Rac1 relocation contribute to endothelial
barrier integrity and that AKAP220 is involved in this process.
Kategorie: Vortrag
Vortrag 33
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: Tracking and quantitative analyses of fluorescence-tagged fusion proteindynamics at endothelial cell junctions
Autoren: Seebach J.(1),Lenk J.(1),Abu Taha A.(1),Schnittler H.(1),
Adressen:(1)Münster|Anatomy and Vascular Biology|Münster|Germany;
email:seebach@uni-muenster.de
Abstract:
Dynamics of cell junction protein-complexes are essential during remodelling in
inflammation, angiogenesis and wound healing but the molecular background is
poorly understood. Recent advances in the developments of fluorescence tagged
protein- technologies and -expression together with fast speed microscopic
techniques such as spinning disc microscopy allows generating time-lapse
movies with high spatial and temporal resolution over long time periods. VEcadherin-mCherry fusion protein was expressed in human umbilical vein
endothelial cells by lentiviral gene transfer. The fusion protein was functionally
active and localized at cell junctions. Time-lapse recording by fast spinning disc
microscopy uncovered high VE-cadherin dynamics. To analyse those fluorescenttagged fusion proteins quantitatively we developed a new software tool. In a first
step the algorithm identifies the outline of all cells within the image. As it
distinguishes between free cell borders and cell junctions, it is possible to follow
junction dynamics in both sparse cell cultures and highly confluent endothelial cell
layers. In a second step we used the segmentation to extract parameters of
interest from the images, such cell perimeters, cell movement or protein
concentration and dynamics in defined regions along the junctions. The modular
concept of the software makes it easy to adapt and expand the analysis to
different problems. Using this tool, we demonstrate a thrombin-dependent
decrease in VE-cadherin dynamics at the cell junctions. This result was confirmed
by fluorescence recovery after photobleaching (FRAP). Thus, the developed
image analyses software allows segmentation and quantitative analyses of
protein dynamics at endothelial cell junctions.
Kategorie: Vortrag
Vortrag 34
Rubrik: 7.Immunbiologie
Abstract Nr.:7
Titel: Tandem peptide blocks pemphigus vulgaris skin blistering in vivo by
preventing loss of desmoglein 3 binding and p38mapk activation
Autoren: Spindler V.(1),Kempf B.(2),Waschke J.(1),
Adressen:(1)Ludwig Maximilians University München|Institute of Anatomy and
Cell Biology, Lehrstuhl I|München|Germany; email:volker.spindler@med.unimuenchen.de; (2)University Würzburg|Institute of Anatomy and Cell Biology,
Lehrstuhl III|Würzburg|Germany
Abstract:
In the life-threatening tissue-specific autoimmune disease pemphigus vulgaris,
epidermal blisters are caused by autoantibodies primarily targeting desmosomal
cadherins desmoglein (Dsg) 3 and Dsg1 leading to loss of keratinocyte cohesion.
Due to limited deeper insights in disease pathogenesis current therapy is
restricted to relatively unspecific long term immunosuppression. Both direct
inhibition of Dsg trans-interaction as well as altered intracellular signaling likely
contribute to loss of cell adhesion. Here, we applied a tandem-peptide (TP) in
vivo consisting of two connected peptide sequences each targeting the Dsg
adhesive interface. TP was capable to block autoantibody-mediated direct
interference of Dsg3 transinteraction as revealed by atomic force microscopy and
optical trapping. Importantly, it was effective to completely abrogate autoantibodymediated skin blistering in mice. Relevant for a possible therapeutic use in
patients, TP was effective also when applied topically. In addition, TP blunted
autoantibody-induced activation of p38MAPK, a central signaling pathway in
pemphigus pathogenesis. These data indicate that p38MAPK links autoantibodyinduced direct inhibition of Dsg binding to skin blistering. By limiting both loss of
Dsg trans-interaction and subsequent p38MAPK activation, TP may serve as
promising treatment option in pemphigus vulgaris.
Kategorie: Vortrag
Vortrag 35
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: Tail-anchored proteins on peroxisomes – common targeting to a third
subcellular compartment?
Autoren: Islinger M.(1),Bonekamp N.(1),Almeida M.(1),Guimaraes S.(1),Castro
I.(1),Camoes F.(1),Schrader M.(1),
Adressen:(1)Universidade de Aveiro|Centre for Cell Biology|Aveiro|Portugal;
email:markus.islinger@ua.pt
Abstract:
Tail-anchored (TA) proteins represent a large group of functionally diverse
proteins which share a single membrane spanning alpha helix close to their Cterminus and fulfil a variety of essential cellular functions. Because the membrane
tail constitutes the only targeting sequence and emerges from the ribosome only
after termination of translation, TA proteins must insert into their target
membranes – endoplasmic reticulum (ER), mitochondria (MITO) and
peroxisomes (PO) – by post-translational mechanisms. Thus, specific targeting
processes have to ensure that the nascent proteins reach the correct destination.
Presently, only proteins contributing to the ER sorting have been characterized in
detail, whereas knowledge on MITO and PO TA protein targeting remains scarce.
PO are essential for human health and development. They fulfil important
metabolic functions in lipid and ROS metabolism influencing e.g. neuronal
development and ageing. Now there is firm evidence that a growing number of TA
proteins localizes to PO including Fis1, Mff or GDAP1, which contribute to PO
(and MITO) division. To unravel if the sharing of TA proteins between both
organelles is a common phenomenon, we screened if more mitochondrial TA
proteins are targeted to PO. Indeed, a subset of proteins showed PO localization,
whereas others were only found on MITO. Furthermore, we identified and
confirmed the existence of TA proteins solely targeted to PO in WT-cells but to
MITO when the formation of PO was blocked in delta-Pex19-cells. Supplemented
with bioinformatics, these findings support our concept of the PO-MITO
connection suggesting their closer mechanistic and functional interrelationship.
Kategorie: Vortrag
Vortrag 36
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: The Cohen syndrome-associated protein COH1 is a novel, giant Golgi
matrix protein required for Golgi integrity
Autoren: Seifert W.(1),Kuehnisch J.(2),Maritzen T.(3),Bachmann S.(1),Horn
D.(2),Haucke V.(3),Hennies H. C.(4),
Adressen:(1)Charité - Universitaetsmedizin Berlin|Institute for Vegetative
Anatomy|Berlin|Germany; email:wenke.seifert@charite.de; (2)Charité Universitaetsmedizin Berlin|Institute of Medical and Human
Genetics|Berlin|Germany; (3)Freie Universitaet and Charité Universitaetsmedizin Berlin|Institute of Chemistry and
Biochemistry|Berlin|Germany; (4)Universitaet zu Koeln|Cologne Center for
Genomics|Cologne|Germany
Abstract:
Mutations in COH1 (VPS13B) are well established to cause autosomal recessive
Cohen syndrome, which is mainly characterized by mental retardation, postnatal
microcephaly, pigmentary retinopathy, and intermittent neutropenia. However, the
biochemical characteristics, cellular localization, or functional role of the encoded
protein COH1 (3997aa) have so far not been addressed. Our cell biological
analysis showed strong co-localization of COH1 with the cis-Golgi marker protein
GM130 which was preserved even upon chemical disruption of the Golgi
architecture. Further biochemical analysis showed that COH1 is a peripheral
membrane protein similar to its remote homologue, Vps13p in yeast. Vps13p has
been found to regulate anterograde and retrograde vesicular transport of
transmembrane proteins between the prevacuolar compartment and the transGolgi network. Accordingly, we found that loss of COH1 upon RNAi impairs the
ability of the Golgi ribbon to (re)assemble and thus induces fragmentation into
mini-stacks. Furthermore, COH1 regulates the formation of Golgi-derived
membrane tubules consistent with its possible function in intracellular membrane
traffic. Human adult skin fibroblasts from Cohen syndrome patients carrying
frameshift or nonsense mutations display a similar disruption of the Golgi complex
which agrees with our observations in HeLa cells. In summary, our study
identifies COH1 as a Golgi matrix protein required for maintaining Golgi integrity
and function. We suggest that our results provide an improved insight into the
molecular function of COH1 in Golgi membrane traffic which in the future may
help to unravel its role in brain development, neuronal function, and general
pathology in patients with Cohen syndrome.
Kategorie: Vortrag
Vortrag 37
Rubrik: 6.Reproduktionsbiologie
Abstract Nr.:6
Titel: Localization of ck2 subunits and its possible binding partners tnp1 and kif5c
in the murine testis
Autoren: Mannowetz N.(1),Kartarius S.(2),Montenarh M.(2),Wennemuth G.(1),
Adressen:(1)Universität des Saarlandes|Institut für Anatomie und
Zellbiologie|Homburg/Saar|Deutschland; email:nadja.mannowetz@uniklinikumsaarland.de; (2)Universität des Saarlandes|Institut für medizinische Biochemie
und Molekularbiologie|Homburg/Saar|Deutschland
Abstract:
Protein kinase CK2 regulates a variety of cellular processes, such as
gametogenesis. The holoenzyme is a heterotetramer consisting of two regulatory
beta-subunits and the two catalytic subunits alpha and alpha'. The individual
subunits have been shown to bind to an abundance of cellular proteins in various
cell types, i. e. the transition nuclear protein 1 (TNP1). It has been demonstrated
that expression of both TNP1 and CK2 peaks in pachytene spermatocytes and
round spermatids. In neurons, CK2alpha' interacts with the neuronal motor
protein KIF5C, which, in turn, has been shown to be expressed also in
spermatids. With regard to a possible interaction between CK2 and TNP1 and
KIF5C respectively, we asked here for the localization of CK2 subunits, TNP1 and
KIF5C
in
spermatogenic
cells
and
mature
spermatozoa.
With
immunohistochemistry we found CK2beta and TNP1 to be present in cells of
early spermatogenesis, whereas CK2alpha, CK2alpha' and KIF5C were localized
in late spermatogenic cells. Immunofluorescence with epididymal sperm showed
CK2alpha-, TNP1- and KIF5C-specific signals in the acrosome, whereas CK2beta
and CK2alpha' were additionally present in the midpiece. Immunogold labeling
revealed a subcellular localization of CK2alpha and KIF5C in the developing
acrosome. However, for CK2beta, CK2alpha' and TNP1 no or only faint signals
restricted to the midpiece were visible. Although we so far did not directly
demonstrate interactions between CK2 proteins and TNP1 and KIF5C
respectively, we conclude that TNP1 and KIF5C are possible binding partners for
CK2 thereby regulating both spermatogenesis and sperm functions.
Kategorie: Vortrag
Vortrag 38
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: Activation of the bumetanide-sensitive na+,k+,2cl- -cotransporter nkcc2 is
facilitated by tamm-horsfall protein in a chloride-sensitive manner
Autoren: Mutig K.(1),Kahl T.(1),Saritas T.(1),Godes M.(2),Rampoldi L.(3),Kumar
S.(4),Bates J.(4),Gamba G.(5),Bachmann S.(1),
Adressen:(1)Charité-Universitätsmedizin Berlin|Institut für
Anatomie|Berlin|Deutschland; email:kerim.mutig@charite.de; (2)CharitéUniversitätsmedizin Berlin|Institut für Physiologie|Berlin|Deutschland; (3)Dulbecco
Telethon Institute|Molecular Genetics of Renal Disorders Unit|Milan|Italy;
(4)University of Oklahoma Health Sciences Center|Department of
Nephrology|Oklahoma City|USA; (5)Instituto Nacional de Ciencias Médicas y
Nutrición Salvador Zubirán|Molecular Physiology Unit|Mexico City|Mexico
Abstract:
Active transport of NaCl in the thick ascending limb (TAL) is accomplished by
Na+,K+,2Cl--cotransporter (NKCC2). The activation of NKCC2 depends on
intracellular chloride concentration (C[i]) and includes its amino-terminal
phosphorylation. We hypothesized that co-expressed Tamm Horsfall protein
(THP) modulates NKCC2 activity in TAL. Effects of THP on NKCC2
phosphorylation (T96/T101) and transport activity were studied in THP-deficient
(THP-/-) and wild type (WT) mice, cultured TAL cells, and frog oocytes. THP-/mice displayed decreased phosphorylation of NKCC2 (-49%, p<0.05) compared
to WT mice. Cultured TAL cells with low endogenous THP levels displayed sharp
increases in NKCC2 phosphorylation (+38%, p<0.05) along with a pronounced
decrease of C[i] (-40.4%, p<0.05) upon transfection with THP. In NKCC2expressing frog oocytes, co-injection with THP cRNA significantly enhanced the
activation of NKCC2 under low chloride hypotonic stress (+112% vs. +235%,
p<0.05). Stimulation of the vasopressin V2 receptor pathway by V2R agonist
(dDAVP; 30 min) resulted in enhanced NKCC2 phosphorylation in WT mice and
cultured TAL cells transfected with THP whereas in the absence of THP, NKCC2
phosphorylation upon dDAVP was blunted in both systems. Attenuated effects of
furosemide along with functional and structural adaptation of the distal convoluted
tubule in THP-/- mice further supported the notion that NaCl reabsorption was
impaired in TAL lacking THP. In summary, these results are compatible with a
permissive role for THP in the modulation of NKCC2-dependent TAL salt
reabsorptive function.
Kategorie: Vortrag
Vortrag 39
Rubrik: 8.Neuroregeneration/Neurodegeneration
Abstract Nr.:8
Titel: Participation of oligodendrocytes in early ms lesion formation
Autoren: Kipp M.(1),Berger K.(2),Amor S.(1),Beyer C.(2),
Adressen:(1)VU University Medical Centre De
Boelelaan|Pathology|Amsterdam|The Netherlands; email:mkipp@ukaachen.de;
(2)RWTH Aachen University|Neuroanatomy|Aachen|Germany
Abstract:
In the last years several studies described clusters of activated microglia cells in
the normal appearing white matter of Multiple sclerosis (MS) patients. This
phenomenon was named “pre-active lesion”.
Oligodendrocyte stress has been linked to pre-active lesion formation. We, thus,
speculate that “stressed” oligodendrocytes are implicated in microglia
accumulation by the secretion of chemokines and thus actively participating in
pre-active lesion formation in MS.
Stimulated OLN93 cells and primary rat oligodendrocyte cultures were analyzed
for chemokine expression and release by rt-PCR and ELISA. Transwell
chemotaxis assays were performed to demonstrate that stimulated
oligodendrocytes attract microglia cells in vitro. Effects of short-term cu-treatment
were analyzed with immunhistochemistry (IHC). Dynamic expression and cellular
distribution of chemokines was analyzed in cuprizone-treated animals by
Affymetrix® microarray, rt-PCR and in situ hybridization (ISH). Loss-of-function
studies were performed to study the role of CXCL10 in microglia accumulation.
Stressed oligodendrocytes expressed and secreted different chemokines, among
CXCL10, paralleled by their ability to attract microglia cells in vitro. Short-term
cuprizone exposure led to heavy oligodendrocytes stress and microglia
accumulation in the absence of demyelination. ISH showed that oligodendrocytes
express CXCL10 mRNA. CXCL10 deficiency led to a significant decrease in the
accumulation of Iba1-positive cells in the cuprizone affected brain compared to
wildtype animals.
We showed that oligodendrocytes are indeed a source of chemokines under
stress conditions and that they are functionally involved in microglia activation.
Further studies have to show the relevance of our findings for MS pre-active
lesion formation and progression.
Kategorie: Vortrag
Vortrag 40
Rubrik: 8. Neuroregeneration/Neurodegeneration
Abstract Nr.:
Titel: Alternative microglia activation in the 6-hydroxydopamine mouse model of
parkinson´s disease
Autoren: Spittau B.(1),Zhou X.(2),Haas S.(3),Duckert M.(3),Wree A.(3),
Krieglstein K.(2),
Adressen:(1)Albert-Ludwigs-Universität|Institut für Anatomie und Zellbiologie,
Abteilung für Molekulare Embryologie|Freiburg|Deutschland;
email:bjoern.spittau@anat.uni-freiburg.de; (2)Albert-Ludwigs-Universität
Freiburg|Institut für Anatomie und Zellbiologie, Abteilung für Molekulare
Embryologie|Freiburg|Deutschland; (3)Universität Rostock|Institut für
Anatomie|Rostock|Deutschland
Abstract:
Microglia activation plays a critical role during progression of several central
nervous system diseases, including Alzheimer´s disease and Parkinson´s
disease. However, recent studies suggest that microglia are able to adopt two
distinct activation modes in vitro and in vivo. Whereas classical microglia
activation is induced by toll-like receptor agonists such as LPS, Interleukin-4 (IL4)
has been shown to promote alternative microglia activation which is characterized
by upregulation of Arginase-1 and YM-1. We have previously demonstrated that
IL4-treated microglia have neuroprotective effects on primary midbrain
dopaminergic (mDA) neurons and that IL4-induced alternative microglia activation
is enhanced by TGF-beta. Here, we address the question whether alternative
microglia activation takes place during the course of the 6-hydroxydopamine (6OHDA) mouse model of Parkinson´s disease in vivo. Injection of 6-OHDA into
the medial forebrain bundle resulted in substantial loss of mDA neurons
accompanied by accumulation of activated microglia in the substantia nigra (SN)
after 4 days. Interestingly, we demonstrate the presence of IL4-positve cells
exclusively in the lesioned SN at the same time. Moreover, TGF-beta1
immunoreactivity was observed in neurons and astrocytes in lesioned and nonlesioned midbrains. Taken together, our results suggest that TGF-beta1 and IL4
might regulate the microglia activation mode during the 6-OHDA lesion to mediate
neuroprotection and recovery.
Kategorie: Vortrag
Vortrag 41
Rubrik: 11.Neuroimmunologie
Abstract Nr.:1
Titel: Cpg oligodeoxynucleotides induce antimicrobial peptide cathelicidin
expression in primary glial cells
Autoren: Brandenburg L.(1),Tauber S.(2),Albrecht L.(1),Jansen S.(1),Wruck
C.(1),Pufe T.(1),
Adressen:(1)RWTH University Aachen|Department of Anatomy and Cell
Biology|Aachen|Germany; email:lbrandenburg@ukaachen.de; (2)RWTH
University Aachen|Neurology|Aachen|Germany
Abstract:
During bacterial infection antimicrobial peptides are synthesized as an important
part of the innate immune system but little is known about their expression and
function in the brain. The aim of this study was to investigate the involvement of
the pattern-recognition-receptor toll-like receptor 9 (TLR9) in the expression of
cathelin-related antimicrobial peptide (CRAMP) in primary glial cells (astrocytes
and microglia).
We examined the expression of CRAMP after treatment with TLR9 agonist
unmethylated cytosine-guanine oligodeoxynucleotide motifs (CpG-DNA) in
primary glial cells using real-time RT-PCR in vitro and in vivo by
intracerebroventricular infusions of CpG-DNA in mice using immunofluorescence.
The pathways which regulate the expression of CRAMP have been identified.
Using a mouse meningitis model, we investigated the CRAMP expression in
TLR9-knock out and wildtype mice using immunofluorescence. Furthermore we
examined for changes in the extracellular signal-regulated kinase (ERK) signal
transduction pathway after stimulation with toll-like receptor 9 ligands in glial and
transfected HEK293 cells.
We show CpG-DNA-induced increase of CRAMP expression by real-time RTPCR and immunofluorescence in glial cells. For expression of CRAMP in glial
cells different signal transduction pathways are involved. Furthermore, we
demonstrated that CpG-DNA-induced ERK1/2 phosphorylation is depended on
TLR9. Additionally, the scavenger receptor MARCO (macrophage receptor with
collagenous structure) is also involved in CpG-DNA-induced cell activation and
CRAMP expression.
Taken together, these results suggest that cathelicidins produced by glial and
other cells play an important part in the innate immune response against
pathogens in central nervous system bacterial infections.
Kategorie: Vortrag
Vortrag 42
Rubrik: 8.Neuroregeneration/Neurodegeneration
Abstract Nr.:8
Titel: Norrin protects photoreceptors from apoptotic cell death
Autoren: Braunger B.(1),Ohlmann A.(1),Koch M.(2),Tanimoto N.(3),Yang
Y.(4),Cvekl A.(4),Boesl M.(5),Seeliger M.(3),Tamm E.(1),
Adressen:(1)Regensburg|Humananatomie und
Embryologie|Regensburg|Deutschland; email:Barbara.Braunger@vkl.uniregensburg.de; (2)Regensburg|Humanantomie und
Embryologie|Regensburg|Deutschland; (3)Tübingen|Institute for Ophthalmic
Research|Tübingen|Deutschland; (4)Albert Einstein College of Medicine|Jack and
Pearl Resnick Campus|New York|USA; (5)Max Planck Gesellschaft|Max Planck
Institute of Neurobiology|Martinsried|Deutschland
Abstract:
Purpose:
Norrin is a signaling molecule, which binds to Fz4 and activates
the canonical Wnt-pathway. We were interested to learn, if overexpression of
norrin in the retina of mice is a mechanism to protect photoreceptors from lightinduced damage, a model of photoreceptor degeneration.
Methods:
Transgenic mice (Rpe65-Norrin) which overexpress norrin in the
retinal pigment epithelium were generated, characterized, and exposed to light
damage. Their phenotype of was analyzed by Western blotting, real time RTPCR, semithin sectioning, immunohistochemistry and electroretinography (ERG).
Wnt-pathway reporter mice were crossbred to analyze activation of the canonical
Wnt-pathway.
Results:
Rpe65-Norrin mice do not express an obvious retinal phenotype.
High amounts of Norrin were detected in their retinae by Western blotting and
immunohistochemistry. Rpe65-Norrin mice showed activation of Wnt-signaling.
30 h after light damage, significantly fewer TUNEL-positive cells were detected in
the outer nuclear layer (ONL) of transgenic mice as compared to wild-type
littermates. 7 and 14 days after light damage, the ONL was significantly thinner in
control littermates than in Rpe65-Norrin mice. Structural changes correlated with
functional changes as detected by ERG. Light damage was enhanced after
blocking Wnt-signaling with Dickkopf-1. Western blotting showed activation of
stat3/phospho-stat3 signaling. By real time RT-PCR, a significant upregulation of
GFAP and endothelin-2 mRNA was observed in retinae of untreated animals,
while light-treated animals showed a significant upregulation of mRNA for BDNF
and LEDGF.
Conclusion:
Norrin protects photoreceptors from light-induced apoptotic cell
death indicating a neuroprotective role of Norrin via activation of the canonical
Wnt-pathway and upregulation of neuroprotective factors.
Kategorie: Vortrag
Vortrag 43
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: Developmental and functional nature of human ips derived motor neurons
Autoren: Liebau S.(1),Linta L.(1),Stockmann M.(1),Böckers T.(1),
Adressen:(1)Ulm|Anatomie und Zellbiologie|Ulm|Deutschland;
email:stefan.liebau@uni-ulm.de
Abstract:
Human induced pluripotent stem cell (hiPS) derived motoneurons represent a
valuable tool to investigate developmental as well as pathophysiological
mechanisms. The developmental processes of human neurons including
maturation, synaptogenesis as well as the establishment of functional contacts to
target cells have been poorly investigated up to now. Especially pathogenetic
studies, however, essentially require a thorough analysis of these aspects in
order to detect, classify and interpret pathophysiological aberrations. Human
motor neurons differentiate about 10 times slower compared to neurons from
mice or rats, however the sequential steps of maturation are essentially identical.
The first neuritis are determined as dendrites and axons after 2 weeks,
electrophysiological properties of motoneurons including spontaneous action
potentials are detectable 3 weeks later. First synaptic contacts are observed after
42-49 days in culture. From then on these contacts continously further mature
even beyond 12 weeks in vitro. Coculturing of human motoneurons with
myotubes resulted in the establishment of functionally active neuromuscular
junctions innervating their natural target cells. Our study provides a thorough
analysis of the differentiation steps of human iPS derived motoneurons and
introduces a model system to study the neuromuscular interplay in vitro. These
data are essential to study pathophysiological phenotypes especially of
motoneuron diseases.
Kategorie: Vortrag
Vortrag 44
Rubrik: 10.Zentrales Nervensystem/Signaltransduktion und Verschaltung
Abstract Nr.:1
Titel: Interplay of reelin and eph signaling in the development of the hippocampus
Autoren: Bock H.(1),Bouché E.(1),Henkemeyer M.(2),Frotscher M.(3),May
P.(1),Herz J.(4),Romero-Ortega M.(5),
Adressen:(1)Universität Freiburg|Zentrum für
Neurowissenschaften|Freiburg|Germany; email:hans.bock@zfn.uni-freiburg.de;
(2)UT Southwestern Medical Center|Department of Developmental
Biology|Dallas|USA; (3)Zentrum für Molekulare Neurobiologie|Institute for
Structural Neurobiology|Hamburg|Germany; (4)UT Southwestern Medical
Center|Department of Molecular Genetics|Dallas|USA; (5)University of Texas at
Arlington|Department of Bioengineering|Arlington|USA
Abstract:
The secreted neuronal signaling molecule Reelin regulates the positioning and
differentiation of postmitotic neurons in the developing brain. Binding of Reelin to
its canonical receptors, ApoER2 and VLDL receptor, induces a signaling cascade
in responsive neurons involving the activation of Src family kinases and tyrosine
phosphorylation of the intracellular apaptor protein Disabled-1. Additional
transmembrane proteins have been reported to bind to Reelin, including integrins
and the amyloid precursor protein, whose biological relevance as Reelin
receptors is less clear. Here, we report that Reelin binds to the extracellular
domain of EphB receptor family members, a group of proteins that interact with
ephrin receptors to regulate various aspects of central nervous system
development, angiogenesis and tumorigenesis.
Specifically, we show that Reelin can activate EphB forward signaling.
Furthermore, mice lacking EphB1 and EphB2 display a circumscribed positioning
defect of hippocampal pyramidal neurons, similar to Reelin-deficient mice, which
depends on the forward signaling capacity of EphB receptors.
Kategorie: Vortrag
Vortrag 45
Rubrik: 10.Zentrales Nervensystem/Signaltransduktion und Verschaltung
Abstract Nr.:1
Titel: Three-dimensional mapping of thalamocortical connectivity in reeler and
wild type mice by directly correlating dti-based fiber tracking and axonal tracing in
vivo
Autoren: Dávid C.(1),Harsan L.(2),Reisert M.(2),Schnell S.(2),Hennig J.(2),von
Elverfeldt D.(3),Staiger J.(4),
Adressen:(1)Semmelweis Universität|Institut für Humanmorphologie und
Entwicklungsbiologie|Budapest|Ungarn; email:david.csaba@med.semmelweisuniv.hu; (2)Universitätsklinikum Freiburg|Medizin Physik, Radiologische
Klinik|Freiburg im Breisgau|Deutschland; (3)Universitätsklinikum Freiburg|Medizin
Physik, Radiologische Klinik|Freiburg im Bresigau|Deutschland; (4)Georg-August
-Universität|Abteilung Neuroanatomie|Göttingen|Deutschland
Abstract:
The complex connectivity of neurons organized in nuclei or areas gives the brain
incomparably powerful information processing capabilities. MRI based diffusion
tensor imaging (DTI) methods are suitable for mapping this connectivity pattern of
the functioning brain in vivo. However, without a methodologically complementary
but independent validation, the results obtained with MRI-DTI in clinical and
preclinical settings have to be interpreted with caution. Here we validated DTIbased fiber tracking methods with classical anatomical tracing in the same
animals of the thalamocortical projection of reeler and wild type mice, which is
one of the key connection concerning sensory pathways. The reeler mouse is an
ideal control group since, due to a to a developmental neuronal migration
disorder, it has altered thalamocortical projection trajectories. Our novel tracking
algorithm was able to predict differences between the two phenotypes, e.g. the
lesser grade of fasciculation in the internal capsule and differences in 3D
structure, which were confirmed later with anatomical tracing of the same
animals. In contrast to previous studies, where the tracks were lost at the border
of the gray and white matter, the projection was followed from the gray matter of
the ventrobasal thalamic nucleus to the cortical gray matter, where the radially
oriented fibers in the wild type and loop-building ones in the reeler could be
distinguished easily. Three different DTI tracking methods (probabilistic,
deterministic and global) were validated with MicroRuby tracing after the
comparison resulted in a highly significant correlation of the maps.
(Supported by DFG: SFB 780, TP C1)
Kategorie: Vortrag
Vortrag 45a
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:
Titel: Unveiling the functional connectivity of individual barrel cortex neurons
using intrinsic signal optical imaging and whole cell recording in wild type and
reeler mice
Autoren: Julien Guy(1), Ilan Lampl(2),, Jochen Staiger(1),
Adressen:(1)Department of Neuroanatomy, Anatomy Center, University
Göttingen, Kreuzbergring 36, 37075 Göttingen, Germany; (2)Department of
Neurobiology, Weizmann Institute of Science, Rehovot 76100, Israel
Abstract:
The barrel cortex of rodents contains a somatotopic representation of the
mystacial whiskers and is a well-established model for the study of tactile
information processing in the cortex. Our goal is to establish the link between
neuronal morphology and physiological sensory response. To this end, we will
characterize the receptive field (RF) size and response properties of individual,
identified neurons within the barrel C2-related column in urethane anesthetized
mice. The skull over the barrel cortex is thinned to translucency in adult mice. The
position of the C2 barrel and its neighbors is mapped using intrinsic signal optical
imaging, which measures changes in light absorbance due to local variations of
blood flow and oxygenation evoked by single whisker stimulation, delivered by a
computer-controlled piezo actuator. After craniotomy, a patch pipette containing
biocytin is lowered into the barrel C2-related column. Once a cell is patched, its
RF properties, adaptation properties and selectivity to the direction of deflection
are determined. The brain is then perfused, sectioned and the recorded neuron is
stained in order to reconstruct its complete morphology using the Neurolucida
software. This combined analysis of functional properties in vivo and detailed
post-hoc morphology allows us to correlate the position of a neuron in a complex
network to its contribution to the network’s function, providing exquisite
knowledge of its functional connectivity. (Supported by DFG:CRC 889-TP C1)
Kategorie: Vortrag
Vortrag 46
Rubrik: 8.Neuroregeneration/Neurodegeneration
Abstract Nr.:
Titel: Targets of the serotonergic system in the rat laterobasal amygdala: studies
on anxiolytic NPY neurons
Autoren: Bonn M.(1),Van Bockstaele E.(2),Schmitt A.(3),Asan E.(1),
Adressen:(1)Würzburg|Institut für Anatomie und
Zellbiologie|Würzburg|Deutschland; email:maria.bonn@uni-wuerzburg.de;
(2)Thomas Jefferson University|Farber Institute for
Neurosciences|Philadelphia|USA; (3)Würzburg|Molekulare Psychiatrie, Klinik für
Psychiatrie, Psychosomatik und Psychotherapie|Würzburg|Deutschland
Abstract:
Serotonin (5-HT) is the neurotransmitter most intimately associated with stress,
anxiety and \"emotionality\". Serotonergic afferents target limbic brain areas like
the laterobasal amygdala (BLA), and 5-HT-dependent modulation of amygdaloid
emotional stimulus processing has been documented in experimental animals
and humans. However, morphological and functional parameters of interactions
between serotonergic afferents and BLA neurons involved in mediating this
modulation have not been comprehensively analyzed, to date. We have shown
that serotonergic afferents form numerous perisomatic appositions on
Neuropeptide Y (NPY)-containing neurons of the rat BLA, an interneuronal
subpopulation, which has been documented to subserve anxiolytic effects. In the
present study, we applied dual immunoelectron microscopy and verified synaptic
contacts and membrane appositions of serotonergic terminals/axons on NPY-ir
somata. NPY-ir terminals formed exclusively symmetric synapses with unlabeled
structures. Applying conventional and a novel variation of double in situ
hybridization (ISH), we showed that NPY mRNA-reactive neurons in the BLA coexpress 5-HT receptor 1A (5-HT1A) and 5-HT2C but lack 5-HT3A mRNA. In
subsequent triple ISH, we documented co-expression of 5-HT1A and 5-HT2C in
some NPY mRNA-reactive neurons in these nuclei. Our findings provide evidence
for a significant and differential impact of serotonergic innervation of NPY neurons
in the rat BLA through 5-HT1A and 5-HT2C. Additionally, they support the
inhibitory nature of NPY-containing elements. Further analyses of interrelations
between monoaminergic afferents and peptidergic interneurons and pyramidal
cells will yield more detailed insights into the emotion-related circuitries in BLA.
Kategorie: Vortrag
Vortrag 47
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:3
Titel: The soluble guanylate cyclase in nerve endings of the dentin-pulp complex
Autoren: Korkmaz Y.(1),Raab W.(1),Schneider K.(1),Behrends S.(2),Bloch
W.(3),Addicks K.(4),
Adressen:(1)Heinrich-Heine-University|Department of Operative Dentistry,
Periodontics and Endodontics|Düsseldorf|Germany; email:yueksel.korkmaz@uniduesseldorf.de; (2)Technical University of Carolo-Wilhelmina at
Braunschweig|Institute for Pharmacology, Toxicology and Clinical
Pharmacy|Braunschweig|Germany; (3)German Sport University|Department of
Molecular and Cellular Sport Medicine|Cologne|Germany; (4)University of
Cologne|Department I of Anatomy|Cologne|Germany
Abstract:
The exact mechanism of the sensory neurotransmission from the dentinoenamel
junction to the terminal nerve endings in the dentin-pulp complex is still not clear.
Despite absence of nerve endings, the dentinoenamel junction is the most
sensitive region of dentin. Therefore, several mechanisms were proposed to
explain the neurotransmission in the dentin-pulp complex including
hydrodynamic, odontoblast receptor and free nerve endings transduction
mechanisms. In heterodimer forms (alpha beta , alpha beta ), soluble
guanylate cyclase (sGC) is activated by nitric oxide (NO) to convert GTP to cGMP
regulating neurotransmission. To test the hypothesis that sGC mediates NOdependent neurotransmission in the dentin-pulp complex, decalcified frozen
sections of rat (n=6) molars were double-stained using NF200 (a marker for
myelinated A-fibers) with antibodies against the alpha1-, beta1-, and alpha2subunit of sGC. The alpha1-, beta1- and alpha2-subunit of sGC were colocalized
with NF200 in nerve fibers and nerve endings in the dentin-pulp complex. From
our earlier results, it is known that NO is generated by nNOS and eNOS in
odontoblasts. Based on these results we propose a new mechanism for
neurotransmission in dentin: odontoblasts with their processes extend to the
dentinoenamel junction mediating NO- as well as NO-cGMP-dependent
neurotransmission from dentinoenamel junction up to nerve endings in the dentinpulp complex.
Kategorie: Vortrag
Vortrag 48
Rubrik: 9.Peripheres und vegetatives Nervensystem
Abstract Nr.:9
Titel: Expression pattern and localization of alpha-synuclein in the human enteric
nervous system
Autoren: Wedel T.(1),Zorenkov D.(2),Deuschl G.(2),Egberts J.(3),Becker
T.(3),Böttner M.(1),
Adressen:(1)Christian-Albrechts-Universität zu Kiel|Anatomisches
Institut|Kiel|Deutschland; email:t.wedel@anat.uni-kiel.de; (2)UKSH Campus
Kiel|Klinik für Neurologie|Kiel|Deutschland; (3)UKSH Campus Kiel|Klinik für
Allgemeine Chirurgie und Thoraxchirurgie|Kiel|Deutschland
Abstract:
Background & aims:
Alpha-synuclein (a-syn) is abundantly expressed in the central nervous system
supposed to be involved in the regulation of neurotransmission. Insoluble fibrils of
phosphorylated a-synuclein (p-a-syn) have been implicated in several
neurodegenerative diseases (e.g. Parkinson disease, Alzheimer disease). The
aim of the study was to determine the gene expression pattern and localization of
a-syn in the human enteric nervous system (ENS).
Material & methods:
Human colonic specimens (n=13, 15-83 years) were processed for a-syn and p-asyn immunohistochemistry. Colocalization of a-syn was assessed by duallabeling with pan-neuronal markers (PGP 9.5, HuC/D). For site-specific gene
expression studies (RT-q-PCR), material was obtained from full-thickness
sections, tunica muscularis, submucosa, mucosa, and isolated enteric ganglia
harvested by laser microdissection (LMD).
Results:
a-syn was expressed in all intestinal layers with highest levels detectable within
the tunica muscularis and submucosa. Ganglia isolated by LMD showed high
expression of a-syn mRNA exceeding the expression of neuron-specific enolase.
Myenteric and submucosal ganglia and nerve fibers were immunoreactive for asyn. Dual-labeling revealed colocalization of a-syn with both neuronal markers. pa-syn immunoreactivity was consistently observed in adult specimens beyond the
5th decade.
Conclusions:
a-syn is abundantly expressed in the human ENS involving all intramural nerve
plexus layers and both neuronal somata and processes. The presence of p-a-syn
within the ENS is a physiological finding in adults with increasing age and may
not be regarded as a pathological correlate. The data provide a basis to evaluate
altered a-syn expression patterns in the ENS of patients with neurodegenerative
diseases.
Kategorie: Vortrag
Vortrag 49
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:3
Titel: Atomoxetine alters leves of synaptic proteins in adult and fetal rat brain
Autoren: Udvardi P.(1),Liebau S.(1),Dreyhaupt J.(2),Fegert J.(3),Böckers
T.(1),Ludolph A.(3),
Adressen:(1)Universität Ulm|Institut für Anatomie und
Zellbiologie|Ulm|Deutschland, Baden-Württemberg; (2)Universität Ulm|Institut für
Epidemiologie und Medizinische Biometrie|Ulm|Deutschland, BadenWürttemberg; (3)Universität Ulm|Klinik für Kinder- und
Jugendpsychiatrie/Psychotherapie|Ulm|Deutschland, Baden-Württemberg
Abstract:
The pathogenesis of psychiatric disorders as well as the mechanisms of the
pharmacological treatments are still not completely understood. Attentiondeficit/hyperactivity disorder (ADHD) is the most frequently diagnosed
neuropsychiatric disorder in childhood. As atomoxetine is the first non-stimulant
compound licensed for the treatment of ADHD, we addressed the issue of its
impact on the developing brain in rodents. Here we demonstrate that acute
atomoxetine treatment (3 mg/kg, i.p.) of pregnant Sprague Dawley rats (E12 E18) significantly shifted expression levels of genes coding for synaptic proteins
solely in the dams brains, whereas synaptic protein levels were altered in both
adults and fetuses. In the dams brain atomoxetine reduced levels of
monoaminergic transporters in prefrontal cortex and striatum (STR). In the adult
STR we furthermore identified significantly increased levels of mRNA and protein
of the vesicular glutamate transporter 1, the presynaptic vesicle marker
synaptophysin (Syp) and the ADHD candidate gene Snap25. Syp levels were
also increased in the mesencephalon (MES) of adults. However, in the fetal brain,
atomoxetine elicited significantly reduced protein levels of N-methly-D-aspartate
receptor subunits (NR1, NR2B) and increased histone 2B protein levels in both
MES and STR. In summary these data clearly demonstrate that atomoxetine’s
cellbiological effects are not limited to the well-characterized inhibition of
norepinephrine reuptake, but also imply development-dependent action on brain
biochemistry. Furthermore our in vivo study strongly indicates, that atomoxetine’s
impact may also alter neuronal and synaptic plasticity.
Kategorie: Vortrag
Vortrag 50
Rubrik: 7.Immunbiologie
Abstract Nr.:7
Titel: An extracellular ndpk amino acid motif is the epitope for conformational
antibodies stimulating the human β1-adrenoceptor
Autoren: Schlipp A.(1),Jahns V.(2),Lohse M.(3),Nikolaev V.(4),Jahns R.(5),
Adressen:(1)Ludwig Maximilians Universität|Anatomie 1|München|Deutschland;
email:angela.schlipp@med.uni-muenchen.de; (2)Universität Würzburg|Institut für
Pharmakologie und Toxikologie|Würzburg|Deutschland; (3)Universtität
Würzburg|Institut für Pharmakologie und Toxikologie|Würzburg|Deutschland;
(4)Georg August Universität Göttingen|Abteilung für Kardiologie und
Pneumologie|Göttingen|Deutschland; (5)Universitätsklinikum
Würzburg|Interdisziplinäre Biomaterial- und Datenbank Würzburg
(IBDW)|Würzburg|Deutschland
Abstract:
Dilated cardiomyopathy represents a major health problem in younger adults and
a subgroup of DCM is associated with autoimmune pathogenic mechanisms
which might be induced by anti-beta1-adrenoceptor autoantibodies (beta1aabs)
stimulating the human fight-or-flight receptor. Diagnostic tests and specific
antibody-targeted therapeutics are still lacking, as detailed knowledge on the
amino acid residues involved in antibody-dependent allosteric regulation of the
beta1-adrenoceptor activity is unavailable. We sought to identify extracellular
protein residues of the second extracellular beta1-adrenoceptor loop (beta1ECII)
essential for the recognition and allosteric adrenoceptor activation by
conformational beta1aabs.
We synthesized cyclic peptides (CP) mimicking the beta1- or beta2-ECII.
Antibodies binding to CPs are neutralized and unavailable for receptor activation.
In the beta1ECII-CP we systematically mutated every non-conserved residue to
alanine and tested the CP-mutants for their capacity to block the functional effects
of disease-causing polyclonal rat, rabbit and monoclonal mouse anti-β1-abs as
well as aabs from immune DCM patients in a cell-based functional assay. This
assay measures cAMP-responses of living cells expressing human beta1-AR and
a fluorescence resonance energy transfer (FRET)-based cAMP sensor. Exposure
to agonists or stimulating beta1-aabs decreases FRET signals. FRET-responses
after incubation of the antibodies with CP-mutants were compared with those of
unblocked antibodies.
Our results indicate that stimulating animal abs as well as aab preparations from
DCM-patients recognize a NDPK amino acid motif within the beta1ECII. These
findings might help to define and diagnose immune DCM, and develop
therapeutic aab-scavenger peptides.
Kategorie: Vortrag
Vortrag 51
Rubrik: 7.Immunbiologie
Abstract Nr.:7
Titel: The soluble ceacam8 - ceacam1 interaction triggers tcr and bcr costimulatory signals in vitro and in vivo
Autoren: Singer B.(1),Scheffrahn I.(2),Klaile E.(3),Schadendorf D.(4),Shively
J.(5),Slevogt H.(3),Öbrink B.(6),Ergün S.(7),
Adressen:(1)Duisburg-Essen|Anatomy/UK-Essen|Essen|Deutschland;
email:bbsinger@gmx.de; (2)Duisburg-Essen|Clinic of Gastroenterology and
Hepatology|Essen|Deutschland; (3)University Hospital Jena|ZIK
Septomics|Jena|Germany; (4)Duisburg-Essen|Department of Dermatology,
Venerology, and Allergology|Essen|Germany; (5)Beckman Research Institute of
the City of Hope|Graduate School of Biological Sciences|Duarte, CA|USA;
(6)Karolinska Institutet|CMB/MCB|Stockholm|Sweden; (7)DuisburgEssen|Anatomy/UK-Essen|Essen|Germany
Abstract:
In general, granulocytes are known as leukocyte subpopulation with the shortest
half-live time. As part of the innate, non-specific immune system, granulocytes
form the first line of defense against anything foreign without conferring longlasting immunity. Because they are not picky about what bacteria or viruses they
attack and what kind of organism directly or indirectly (e.g. via cytokines) regulate
them, granulocytes were believed to represent a quite primitive type of immune
cells. Thus, they were thought of as terminally differentiated without considerable
de novo synthesis of proteins. However, recent progress has shown, that
granulocytes are able to synthesize pro-inflammatory cytokines [e.g., TNF, IL-1,
IL-8, MIP-1] and angiogenic factors [e.g. VEGF] in response to certain stimuli.
Here we show that the de novo synthesize of soluble CEACAM8 (sCEACAM8)
was provoked by GM-CSF/G-CSF treatment as well. We also detected an
increased level of sCEACAM8 in granulocytes isolated from patients suffering
psoriasis reflecting the immune cell priming in vivo. Moreover, we identified
CEACAM1 as potent receptor for sCEACAM8. In lymphocytes the sCEACAM8–
CEACAM1 interaction functions as positive co-receptor system of the T-cell
receptor and B-cell receptor in vitro and in vivo. Thus, granulocytes are able to
employ the release of sCEACAM8 to manipulate the function of T and Blymphocytes and presumably further CEACAM1 expressing cell types like
activated endothelial and epithelial cells.
Kategorie: Vortrag
Vortrag 52
Rubrik: 7.Immunbiologie
Abstract Nr.:7
Titel: Expression and regulation of host defense peptide psoriasin at the ocular
surface and in the lacrimal apparatus
Autoren: Garreis F.(1),Gottschalt M.(2),Gläser R.(3),Paulsen F.(4),
Adressen:(1)Friedrich-Alexander-Universität Erlangen-Nürnberg|Institut für
Anatomie LHII|Erlangen|Deutschland; email:fabian.garreis@anatomie2.med.unierlangen.de; (2)Martin-Luther Universität Halle-Wittenberg|Institut für Anatomie
und Zellbiologie|Halle|Deutschland; (3)Christian-Albrechts
Universität|Universitäts-Hautklinik|Kiel|Deutschland; (4)Friedrich-AlexanderUniversität Erlangen-Nürnberg|Institut für Anatomie LH II|Erlangen|Deutschland
Abstract:
The S100 peptide Psoriasin (S100A7c) has recently been identified as an
antimicrobial peptide of healthy skin and mucosa. The purpose of this study was
to investigate the expression and antimicrobial role of psoriasin at the ocular
surface and in the lacrimal apparatus.
Different tissues of the lacrimal apparatus and ocular surface were systematically
analyzed by means of RT-PCR, Western blot and immunohistochemistry for their
ability to express and produce psoriasin. The inducibility and regulation of
psoriasin were studied in cultivated human corneal as well as conjunctival
epithelial cells after challenge with different pathogen-associated molecular
patterns (PAMPs) from frequent ocular pathogens and proinflammatory cytokines.
Real-time RT-PCR was performed to evaluate the expression and induction of
psoriasin. In addition, tear fluid obtained from different healthy volunteers was
examined by ELISA for its psoriasin concentration.RT-PCR and Western blot
analyses revealed a constitutive expression of psoriasin in cornea, conjunctiva,
nasolacrimal ducts and lacrimal gland. Immunohistochemistry showed strong
staining of Meibomian glands for psoriasin. No induction of psoriasin was
observed after stimulation with supernatants of E. coli whereas supernatants of
Staphylococcus aureus and Haemophilus influenzae significantly increased the
psoriasin mRNA expression. Stimulation with IL-1β and VEGF also strongly
increased the psoriasin transcription. The highest amounts of psoriasin protein
were detected in tear fluid (~170 ng/ml) of healthy volunteers. In conclusion, the
results suggest that psoriasin is produced by the structures of the ocular surface
and is part of the innate immune responses at the ocular surface and the lacrimal
apparatus.
Kategorie: Vortrag
Vortrag 53
Rubrik: 7.Immunbiologie
Abstract Nr.:7
Titel : Anti-tnf alpha therapy alone prevents diabetes before onset and in
combination with anti-cd3 cures diabetes after onset in the iddm rat
Autoren: Jörns A.(1),Ertekin Ü.(1),Meyer zu Vilsendorf A.(2),Lenzen S.(1),
Adressen:(1)Medizinische Hochschule Hannover|Institut für Klinische
Biochemie|Hannover|Deutschland; email:joerns.anne@mh-hannover.de;
(2)Medizinische Hochschule Hannover|Allgemein-, Viszeral- und
Transplantationschirurgie|Hannover|Deutschland
Abstract:
The pro-inflammatory cytokine TNF-alpha was expressed in the immune cell
infiltrate during and after diabetes manifestation. Thus in other autoimmune
diseases the antibody anti-TNF-alpha was a well-established immunomodulatory
agent, but only occasionally used in type 1 diabetes.
Therefore, IDDM (LEW.1AR1-iddm) rats were treated either with anti-TNF-alpha
(5 mg/kg b. wt.) alone or in combination with CD3-AB (0.5 mg/kg b. wt.)
consecutively over 5 days either starting at day 45 before diabetes onset or
immediately after diabetes manifestation. Metabolic parameters in serum samples
were compared with the islet morphology using sequential pancreatic biopsies at
the potential time point of diabetes, at the end of therapy, and 60 days thereafter.
Using the monotherapy of anti-TNF-alpha before diabetes onset the immune cell
migration into the pancreas was completely prevented without apoptotic beta cell
loss in the islets and with a normal high C-peptide concentration. Using anti-TNFalpha alone, however, after diabetes manifestation the apoptotic beta cell death
could not be stopped, only by the combined therapy. The protected and increased
beta cell mass was accompanied with a preservation of the C-peptide
concentration in 60 % of the treated animals, also 60 days after the end of
therapy. Beta cell proliferation rate remained two times higher than the apoptosis
rate, but remained increased compared to the control. In the islets some
remaining immune cells were observed.
After diabetes manifestation only a combined therapy of anti-TNF-alpha and antiCD3 can cure the diabetes whereas before diabetes manifestation anti-TNFalpha protects against immune cell migration into the pancreas.
Kategorie: Vortrag
Vortrag 54
Rubrik: 7.Immunbiologie
Abstract Nr.:7
Titel: Tertiary lymphoid organ development facilitates determinant spreading of
the mp4-specific t cell response
Autoren: Kuerten S.(1),Kerkloh C.(1),Schickel A.(1),Pochmann J.(1),Addicks
K.(1),Lehmann P.(2),Ruddle N.(3),
Adressen:(1)Köln|Institut I für Anatomie|Köln|Deutschland;
email:stefanie.kuerten@uk-koeln.de; (2)Case Western Reserve
University|Department of Pathology|Cleveland|USA; (3)Yale
University|Epidemiology and Public Health|New Haven|USA
Abstract:
Accumulating evidence indicates that in multiple sclerosis (MS), similar to other
autoimmune diseases, B cells aggregate into tertiary lymphoid tissue-like organs
(TLOs) in the target tissue. The characterization of such structures as a niche for
the generation of plasma cells and the activation of T cells independent from the
immune periphery reinforces that the concept of B cell-mediated immunity has to
be critically reevaluated. In patients with secondary-progressive MS the presence
of ectopic B cell follicles has been linked to a younger age at disease onset,
irreversible disability and more pronounced demyelination and loss of neurites in
the cerebral cortex. Here we demonstrate the formation of TLOs in myelin basic
protein (MBP)-proteolipid protein (PLP) fusion protein MP4-induced experimental
autoimmune encephalomyelitis (EAE) of C57BL/6 mice. TLOs were characteristic
of chronic disease, emerging around 40 days after EAE onset and were most
prevalent in the cerebellum, while also being present in the spinal cord and
cerebrum, but absent in the brainstem. TLOs were characterized by B and T cell
compartmentalization, presence of high endothelial venules, lymphatic vessels
and germinal center formation. Furthermore, results obtained by ELISPOT
analysis of the antigen-specific T cell response suggest that TLOs may facilitate
determinant spreading of the MP4-specific T cell response to myelin
oligodendrocyte glycoprotein (MOG). Thus, our data support the notion that TLO
formation can contribute to disease chronification in the context of autoimmunity
and blockade of TLO development may be a therapeutic strategy that needs to be
considered carefully in future studies.
Kategorie: Vortrag
Poster 1
Rubrik: 1.Methoden/Unterricht
Abstract Nr.:
Titel: A gender perspective on the evolution of consciousness: eleven
commandments for the advancement of medical enlightenment.
Autoren: Kurz H.(1),
Adressen:(1)Paracelsus Private Medical University|Tissue Dynamics
Lab|Salzburg|Austria; email:haymo.kurz@email.de
Abstract:
Professors of anatomy, much like the vast majority of god-kings, prophets,
priests, philosophers or gurus in history, used to be recruited - until a few
decades ago - exclusively among males. With the exception of Anna Manzolini
(Bologna, 1714-1774), women were never seen among paid faculty until 200
years later. The questions arise 1) how Homo sapiens males could possibly
achieve the \"remarkable\" feat of restricting sapientia largely to those individuals
that contained only one X-chromosome; 2) whether molecular and cultural
evolution of human brain functions are interdependent; 3) which consequences
may arise from cognitive, moral and reproductive repression vs. emancipation of
women in all fields of science, humanities and arts. Possible answers are
proposed here in the formal disguise of Eleven Commandments. In this attempt, a
key role is assigned to teaching the concept of evolutionary developmental
biology (Evo-Devo) which has gained impetus since completion of the human
genome project 10 years ago. Of note, gene-centered approaches provided
insight how much the genetic material, its expression and change depend on their
interaction with other tissue or environmental factors, thus leading to Eco-Devo. It
is argued here that ecology should also include cultural evolution which acts back
on molecular evolution. This view is substantiated by demonstrating how minute
alterations of DNA may lead to dramatic changes of neocortical growth, how the
human X-chromosome achieved a high density of genes relevant for cognition,
and how consciousness-dependent medical interventions may feed back into
evolutionary changes.
Kategorie: Poster
Poster 2
Rubrik: 1.Methoden/Unterricht
Abstract Nr.:
Titel: Virtual microscopy extended to electron microscopy
Autoren: Jaegermann A.(1),Zanger K.(1),Rohbeck B.(1),Filler T.(1),
Adressen:(1)Universitaet Duesseldorf|Zentrum fuer Anatomie und
Hirnforschung|Duesseldorf|Deutschland; email:andreas.jaegermann@uniduesseldorf.de
Abstract:
When dealing with conventional light microscopy the application of virtual
microscopes is broadly accepted and willingly used to support and extend
curricular lessons of microscopic anatomy in many places. Regarding the electron
microscopy a comparable situation cannot be found, although mostly identical
teaching methods apply. This field of application is still dominated by more or less
completely labelled image atlases, which may be (partially) available for noninteractive web access, too.
With our approach we simplify the integration of high-resolution EM images into a
virtual microscopy environment by linking these images to common virtual slides.
For this purpose matrix-like images from the electron microscope are merged into
one single image which will serve as the input data for our virtual microscope and
will be observable in different magnification steps after some automated
preparation. As with the virtual light microscope slides students will be able to
annotate the electron-optical images. The best of these labels can be added to a
global collection and thereby be made available to all students through a
cascaded system of publishing rights.
Depending on the number of EM images used in curricular or optional lessons, a
few thousands of labels will be created by the students each year. Perspectively
this will lead to a completely labelled, interactive electron-optical collection of
images with a direct connection to light-microscopy contents. An upcoming
evaluation will show the expected increase in recognition capacity of EM images
by the students using this system.
Kategorie: Poster
Poster 3
Rubrik: 1.Methoden/Unterricht
Abstract Nr.:
Titel: Leistungscheck lc uni ulm - a subject-specific progress test Autoren: Fassnacht U.(1),Schröder L.(1),Hanß F.(1),Böckers A.(1),Böckers T.(1),
Adressen:(1)Universität Ulm|Institut für Anatomie und
Zellbiologie|Ulm|Deutschland
Abstract:
Since winter term 2009/2010 the subject-specific progress test of Ulm University
“LeistungsCheck Uni Ulm” measures the momentary anatomical knowledge of
dental students. The survey analyzes the individual development of anatomical
knowledge and identifies influencing factors.
The LC Uni Ulm for all preclinical semesters of dental medicine takes place in the
beginning of the semester consisting of 100 anatomical statements. We use the
true-false question format with a “don’t know” option. The level of difficulty of the
questions has been estimated by the modified Angoff’s method. The test is based
on a weighted blueprint. The test value is the sum of right (1 pt), wrong (-1 pt)
assessments and abstentions (0 pt). After every test run an individual result
report is provided for the participants including individual test value, individual test
value compared with average test value of the semester, individual test value
compared with average test value oft the exam’s semester, and individual test
value for anatomical subdisciplines.
In addition, general information on personal data, previous job training and
education, experience with self-assessment and setting to the LC were obtained.
The results show that the anatomical knowledge of the participants develops nonlinear and corresponds well to the self approximated competence. A significant
increase of anatomical knowledge can only be detected during periods of study
with relevant assessments. Native speakers and students with previous job
training have a small advantage. The LC Ulm provides a reliable feedback for the
students with the chance to adapt the individual learning behavior. Besides,
learning effects can be examined in the field to optimize teaching supply.
Kategorie: Poster
Poster 4
Rubrik: 2. Klinische Anatomie/Makroskopie
Abstract Nr.:2
Titel: Study concerning the morpho-functional parameters of the scapulohumeral
joint ligaments
Autoren: MOTOC A.(1),HOGEA B.(1),STANA L.(1),FOLESCU R.(1),MOISE
M.(2),PETRESCU C.(2),SELARU M.(2),SISU A.(2),
Adressen:(1)UNIVERSITY OF MEDICINE AND PHARMACY “VICTOR
BABES”|DEPARTMENT OF ANATOMY AND
EMBRYOLOGY|TIMISOARA|ROMANIA; email:amotoc@umft.ro; (2)UNIVERSITY
OF MEDICINE AND PHARMACY "VICTOR BABES"|DEPARTMENT OF
ANATOMY AND EMBRYOLOGY|TIMISOARA|ROMANIA
Abstract:
Macroscopic dissection is performed for this study, using the delto pectoral
approach and cutting the following muscles: deltoid, pectoralis major, coraco
brachialis, biceps brachialis (short head), latissimus dorsi, teres major. It have
been performed the following measurements: coraco humeral ligament, inferior
gleno humeral ligament, apparent diameter of the humeral head, greater axis of
the humeral head, greater and smaller axes of the glenoid cavity, and the
distance between the two humeral insertions of the gleno humeral ligament and
inferior coraco humeral ligament, the distance between the two glenoidal
insertions of the two ligaments. Were successfully dissected the scapulo humeral
joints of the 20 formalized bodies, 10 females and 10 males. The conclusion was
that there was no difference between the size of the parameters of the right and
left upper limbs. For each parameter there is an average value between the
minimum and maximum. The coraco humeral ligament has an average length
right / left = 3.8/4.0 cm; the inferior gleno humeral ligament has an average length
right / left = 5.1/5.3 cm; the humeral insertion has an average size right / left of
5.4/5.6 cm; the glenoidal insertion has an average size right / left = 4.5/4.7 cm.
The study highlights a set of intercorrelated average values.
Keywords: ligament, upper limb, scapulo humeral joint.
Kategorie: Poster
Poster 5
Rubrik: 2.Klinische Anatomie/Makroskopie
Abstract Nr.:2
Titel: Friedrich Reinke (1862-1919) and his research on human testicles – the
reinke´s crystals
Autoren: Dräger D.(1),Wree A.(1),
Adressen:(1)Universität Rostock|Institut für Anatomie|Rostock|Deutschland;
email:draeger.desiree@googlemail.com
Abstract:
Purpose: On the occasion of the approaching 150th birthday of Friedrich Reinke
on April 11th 2012 it is an appropriate time to describe some of his anatomical
research results and discuss them anew. Besides the characterisation of Reinke’s
space in the larynx, he described the histological structure of the testes. Reinke’s
crystals are normal constituents of Leydig cells, but their nature and function are
poorly understood. In this study we will examine the Reinke´s crystals of the
Leydig cells whose discovery and significance for the human should be
discussed.
Results: In „Beiträge zur Histologie des Menschen. Teil I. Ueber
Krystalloidbildungen in den interstitiellen Zellen des menschlichen Hodens“
(1896) Reinke described rectangular crystalloide inclusions in the Leydig cells. He
reported that the crystals were found in the cytoplasm of interstitial cells of all
actively spermatogenic testes. By in depth-investigations, Reinke has come to
determine that the crystalloids are proteins which are related with the
spermatogenesis. The function was still unclear to him. Since 1956, Reinke's
crystals have been investigated by electron microscopy and have been noted that
they are polygonal in shape.
Conclusion: Reinke´s crystals are often detectable in adult Leydig cells. These
crystalloids are globular protein subunits, whose functional importance is not
precisely known. As they decrease in number with age, the presence of
degenerative processes is indicated. In addition, they are considered to be
pathognomic in Leydig cell tumour of the testes as well as the ovaries.
Kategorie: Poster
Poster 6
Rubrik: 2.Klinische Anatomie/Makroskopie
Abstract Nr.:2
Titel: Biometric study of heart arterial orifices
Autoren: FOLESCU R.(1),PETRESCU C.(1),MOTOC A.(1),POP E.(1),SISU A.(1),
Adressen:(1)UNIVERSITY OF MEDICINE AND PHARMACY “VICTOR
BABES”|DEPARTMENT OF ANATOMY AND
EMBRYOLOGY|TIMISOARA|ROMANIA; email:roxanafolescu@yahoo.com
Abstract:
A biometric study of heart arterial orifices was carried out on a total of 54 normal
and 15 pathological hearts with coronary atherosclerosis. The 54 normal hearts
were divided by age and sex and the 15 hearts with coronary atherosclerosis
pathology were grouped according to the dynamics of coronary atherosclerosis
lesions: stage I, four hearts, stage II, five hearts, and stage III six hearts. The
circumference of the heart arterial orifices was determined by measuring the
aortic and pulmonary orifice diameters, using a caliper, and afterwards the actual
diameter of the orifice was determined mathematically. The results of the
measurements indicate that the average circumference of the orifice of the
pulmonary artery, in a male adult is 67.9 mm. In females, it is 66.8 mm and in
males it is 69.1 mm. The average of 67.9 mm is between two limits, a minimum of
57 mm and a maximum of 78 mm. In normal adult aortic arterial orifice
circumference is approximately 68.4 mm, with a value of 67 mm, in females and
69.8 mm in males. The average total value is 68.4 mm, being between two limits,
a minimum of 55 mm and a maximum of 81 mm. Pulmonary artery orifice
circumference in pathological hearts is 1.1561 times higher than in normal hearts.
Aortic artery orifice circumference in pathological hearts is 1.1330 times higher
than in normal hearts.
Keywords: normal heart, aortic artery orifice, pulmonary artery orifice.
Kategorie: Poster
Poster 7
Rubrik: 2.Klinische Anatomie/Makroskopie
Abstract Nr.:2
Titel: The risk of injury to the posterior tibial artery in the posterolateral approach
to the tibia plateau. a cadaver study.
Autoren: Clement H.(1),Heidari N.(2),Lidder S.(3),Grechenig S.(4),Tesch
N.(5),Weinberg A.(6),Pichler W.(4),
Adressen:(1)Medical University of Graz|Department of
Traumatology|Graz|Österreich; (2)Royal London Hospital|Department of Trauma
and Orthopaedics|London|UK; email:n.heidari@gmail.com; (3)The Royal London
Hospital|Department of Trauma and Orthopaedics|London|UK; (4)Med.Univ.
Graz|Department of Traumatology|Graz|Österreich; (5)Med.Univ.Graz|Institute of
Anatomy|Graz|Österreich; (6)Med. Univ. Graz|Department of Pediatric and
Adolescent Surgery|Graz|Österreich
Abstract:
Posterolateral tibial plateau fractures are rare and often require posterolateral
buttress plating The purpose of this anatomical study was to provide accurate
data about the inferior limit of dissection by providing measurements from the
lateral joint line to the anterior tibial artery as it pierces the interosseous
membrane. Forty unpaired cadaver adult lower limbs were used for this study.
The anterior tibial artery was identified as it coursed through the interosseous
membrane. The perpendicular distance from the lateral joint line and fibula head
to this landmark was measured. The anterior tibial artery coursed through the
interosseous membrane at 46.3 ± 9.0 mm (range 27 – 62 mm) distal to the lateral
tibial plateau and 35.7 ± 9.0 mm (range 17 – 50 mm) distal to the fibula head.
There was no significant difference between right or left sided knees. (anterior
tibial artery distal to lateral tibial plateau; right : 44.8 ± 8.5 mm, left : 47.8 ± 9.3
mm, and distal to fibula head; right: 34.4 ± 8.6 mm, left: 37.0 ± 9.3 mm).Displaced
posterorlateral tibial plateau fractures require anatomical reduction and
stabilization with a buttress plate. This can only be achieved by gaining access to
the posterolateral tibial cortex. The distal limit of this dissection can be as little as
27 mm from the lateral tibial plateau.The surgeon can expect to be able to use a
small fragment buttress plate of up to 45 mm as this is the mean.
Kategorie: Poster
Poster 8
Rubrik: 2.Klinische Anatomie/Makroskopie
Abstract Nr.:2
Titel: Dynamic hip screw and the risk of injuries of vessels
Autoren: Tesch N.(1),Neubauer T.(2),Grechenig S.(3),Clement H.(3),Tanzer
K.(4),
Adressen:(1)Medizinische Universität Graz|Institut für Anatomie|Graz|Österreich;
email:norbert.tesch@medunigraz.at; (2)Wilhelminenspital|Abt. für
Unfallchirurgie|Wien|Österreich; (3)Med. Univ. Graz|Universitätsklinik für
Unfallchirurgie|Graz|Österreich; (4)Med.Univ. Graz|Universitätsklinik für
Unfalchirurgie|Graz|Österreich
Abstract:
More than 70% of all fractures of the proximal third of the femur are lateral or
peritrochanteric ones. Especially in those cases a high load stability is important.
Therefore the use of a dynamic hip screw and a sliding-hole plate has proved to
be a success. While attaching and fixing the plate, the surgeon has to be aware
of the layout of the vessels on the opposite side of the femur to avoid damaging
them.
With an image intensifier giving a lateral view we inserted dynamic hip screw
plates with 4 holes (produced by Synthes) into 10 cadavers and fixed them with 4
screws each. We then proceeded to dissect the thighs from medially layer by
layer to document injuries of the large vessels of the thigh caused by the fixing
screws.
Of the 40 screws we inserted 7 hit a vessel directly, with the femoral artery being
hit 4 times, the deep artery of the thigh as well as the perforating arteries 1 and 2
being hit once each.
The surgeon has to take special care when inserting and fixing the dynamic hip
screw plate to the femur, because otherwise the proximity of the fixing screws to
the great vessels of the thigh may be the cause of grave injuries to those vessels.
Kategorie: Poster
Poster 9
Rubrik: 2.Klinische Anatomie/Makroskopie
Abstract Nr.:2
Titel: Computerbased analysis of the optimal pin positions for an external fixator
in the area of the ala of ilium
Autoren: Tanzer K.(1),Puchwein P.(1),Pichler W.(1),Tesch N.(2),Clement
H.(1),Nenning T.(1),
Adressen:(1)Med.Univ. Graz|Department of Traumatology|Graz|Österreich;
email:karin.tanzer@medunigraz.at; (2)Med.Univ. Graz|Institute of
Anatomy|Graz|Österreich
Abstract:
Pelvic external fixation is an important technique for primary and definite
treatment of
pelvic ring fractures. The correct placement of the pins in the bone is essential for
the
quality of the stabilization of the pelvic ring. The aim of this diploma thesis was to
analyze the vertical angle to the median-sagittal-plane that is necessary when
inserting a pin for external fixation in the anterior third of the iliac crest (3 cm
posterior to the anterior superior iliac spine) and how long the intraosseous part of
the pin should be when the head of the screw comes to lie in the compact bone of
the Acetabulum without perforating the cartilage.
This information should alleviate the correct placement of the pins when installing
an antero-superior external fixator of the pelvis.
Measurements were performed on 49 computer-tomography images of pelvic
bones under use of software to reconstruct the images three-dimensionally.
Distinction was drawn between female and male pelves. The analyses showed an
average intraosseous length of the pin of 99.90 mm (96.30 mm for female and
101.20 mm for male pelves) and an average angle of 30.30° from vertical (29,60°
for female and 30,50° for male pelves).
When it comes to necessary pin-length, significant differences could be found
between male and female pelves. The resulting angles did not show any
significant differences between male and female pelves.
Up to now there are no papers on this issue where a comparable method is used
to take measurements of the pelvic bone or which analyse the differences in the
found parameters between female and male pelvic bones.
Kategorie: Poster
Poster 10
Rubrik: 2.Klinische Anatomie/Makroskopie
Abstract Nr.:2
Titel: The risk of nerve injury with minimally invasive plate osteosynthesis of distal
fibula fractures: an anatomic study.
Autoren: Neubauer T.(1),Heidari N.(2),Weinberg A.(3),Grechenig S.(4),Tesch
N.(5),Pichler W.(4),Wagner M.(6),
Adressen:(1)Landesklinikum Waldviertel Horn|Abt. für
Unfallchirurgie|Horn|Österreich; email:unfall@horn.lknoe.at; (2)The Royal London
Hospital|Department of Trauma and Orthopaedics|London|UK; (3)Med.Univ.
Graz|Department of Paediatric and Adolescent Surgery|Graz|Austria;
(4)Med.Univ. Graz|Department of Traumatology|Graz|Austria; (5)Med.Univ.
Graz|Institute of Anatomy|Graz|Austria;
(6)Wilhelminenspital|Unfallchirurgie|Wien|Österreich
Abstract:
The aim of our study was to identify the structures which may be at risk of injury
when using a minimally invasive technique for the osteosynthesis of the lateral
malleolus and the influence of the size of the implant on the frequency of injury to
these structures.Forty plates were percutaneously inserted in 20 cadaveric legs.
The region around the plate was then dissected to examine the relation of nerves
and soft tissues to the plate.The superficial peroneal nerve was in direct contact
with the plate in 11 of the 20 cases (55%) of the 10 hole plates. We encountered
only one case of the superficial peroneal nerve skirting the proximal edge of a 6
hole plate (p = 0.0164).Consequently we recommend meticulous attention is paid
to the dissection of soft tissues in both the proximal and distal incisions. The
length of the plate may be checked with intraoperative imaging prior to its
insertion, and the site of both proximal and distal incisions may be marked on the
skin. After careful dissection down to the bone, preserving nerves and tendons,
the periosteal elevator should be introduced both from the proximal as well as the
distal incisions to prepare the extra-periosteal tunnel for the insertion of the plate,
in order to avoid the entanglement of the superficial peroneal nerve with the metal
work, particularly in plates of longer than six holes.
Kategorie: Poster
Poster 11
Rubrik: 2.Klinische Anatomie/Makroskopie
Abstract Nr.:2
Titel: Anatomic studies concerning the sinoatrial artery
Autoren: PETRESCU C.(1),MOTOC A.(1),POP E.(1),FOLESCU R.(2),STANA
L.(2),SELARU M.(2),CIPU D.(3),SISU A.(2),
Adressen:(1)UNIVERSITY OF MEDICINE AND PHARMACY „VICTOR
BABES”|DEPARTMENT OF ANATOMY AND
EMBRYOLOGY|TIMISOARA|ROMANIA; email:codruta.petrescu@gmail.com;
(2)UNIVERSITY OF MEDICINE AND PHARMACY "VICTOR
BABES"|DEPARTMENT OF ANATOMY AND
EMBRYOLOGY|TIMISOARA|ROMANIA; (3)UNIVERSITY OF MEDICINE AND
PHARMACY "VICTOR BABES"|DEPARTMENT OF RADIOLOGY AND MEDICAL
IMAGING|TIMISOARA|ROMANIA
Abstract:
The sinoatrial node is the physiological pacemaker of the heart. Its arterial source
or sources of pressure are therefore essential to be recognized and protected
during surgery at this level. The final anatomic macroscopic study on sinoatrial
node artery (SANA) has been made on a group of 50 adult human hearts. 62% of
specimens (31 specimens) right coronary artery originated from SANA (SANA
right). 16% of specimens (8 hearts) had a double reliance of the sinoatrial node,
arising from the right coronary artery and left SANA, SANA being a branch from
the circumflex artery. 22% of specimens (11 hearts) had the left SANA as
originating from the circumflex artery. A specimen with double right and left SANA
was presenting the left SANA as starting as a posterior left atrial branch and its
general morphology to define such an artery in the sinoatrial node “S” artery.
From the hearts that had the right SANA, it appeared as anterior right atrial
branch in 28 specimens (56%) or right marginal atrial branch in 3 specimens
(6%). SAN right anterior artery starts as ascending branch from the right coronary
artery an average at to 2 cm to its origin, while respecting the first inch of leaving
with downward trajectory the right conal artery. It has been found this option most
frequently in our specimens. In conclusion, anatomic study of sinoatrial node
artery highlights the frequency variations of this artery which must be considered
during surgery at this level.
KEYWORDS: sinoatrial node, epicardium, sinoatrial artery
Kategorie: Poster
Poster 12
Rubrik: 2.Klinische Anatomie/Makroskopie
Abstract Nr.: 2
Titel: Chilaiditi syndrome: anatomical and clinical findings
Autoren: Nimtschke U.(1),Lenz M.(2),Kindler M.(3),Schilling M.(2),Pollak
T.(4),Becker M.(2),Böhnisch M.(1),Schwab W.(1),
Adressen:(1)TU Dresden|Institut für Anatomie|Dresden|Deutschland;
(2)Weißeritztal-Kliniken|Klinik für Allgemein- und
Viszeralchirurgie|Freital|Deutschland; (3)Weißeritztalkliniken|Klinik für Allgemeinund Virzeralchirurgie|Freital|Deutschland; (4)Gemeinschaftspraxis Diagnostische
Radiologie Freital und Dippoldiswalde|Gemeinschaftspraxis Diagnostische
Radiologie Freital und Dippoldiswalde|Freital|Deutschland;
email:wolfgang.schwab@tu-dresden.de
Abstract:
The abnormal hepatodiaphragmatic interposition of the colon was described in
detail by the greek radiologist Dimitrious Cilaiditi in 1919.The so-called Chilaiditi
sign is usually asymptomatic and found as an incidental radiographic sign with a
prevalence rate from 0.025 to 0.28 %. When associated with clinical symptoms
such as abdominal pain or bowel obstruction, it is termed as Chilaiditi syndrome.
In the present study, we present the position of the transverse colon and the right
colon flexur between the left hepatic lobe and the diaphragm that corresponds to
the signs of Chilaiditi and that we found in the cadaver of an 81 year old woman
during routine dissection in the anatomical course. The hepatodiaphragmatic
position of the transverse colon was associated with herniated gastric fundus and
body within a paraesophageal hernia. Furthermore, we describe
computertomographic and intraoperative findings in a case of an 80-year-old
female patient with Chilaiditi syndrome.
The postulated mechanisms in the etiology of Chilaiditi signs include hypoplasia
of liver volume, intestinal malrotation or abnormal motility, laxity of suspendory
ligaments of the liver and paralysis of the phrenic nerve. In the case of the
presented cadaveric study, the connection between the herniated stomach and
the transverse colon by the gastrocolic ligament could contribute to the intestinal
dislocation.
Kategorie: Poster
Poster 13
Rubrik: 2.Klinische Anatomie/Makroskopie
Abstract Nr.: 2
Titel: Morphological individual elements of the accesory and duplicated
saphenous veins
Autoren: STANA L.(1),MOTOC A.(1),PETRESCU C.(1),CEBZAN C.(1),SISU
A.(1),
Adressen:(1)UNIVERSITY OF MEDICINE AND PHARMACY „VICTOR
BABES”|DEPARTMENT OF ANATOMY AND
EMBRYOLOGY|TIMISOARA|ROMANIA;
email:loredana.gabriela.stana@gmail.com
Abstract:
In a group of 60 studied specimens, we identified the presence of the following
accessory saphenous veins: anterior accessory saphenous vein, posterior
accessory saphenous vein, superficial accessory saphenous vein, Leonardo’s
vein and, in addition, saphenous duplication. We considered that both, the
duplicate and the accessory saphenous veins are intermediate position in the
saphenous compartment, unlike the epifascial tributaries or communicating veins.
Of the total group under study, 34 lower limbs (56.67%) showed no venous
elements that can be defined as accessory or duplicated saphenous veins,
according to the set forth criteria. Only 26 specimens (43.33%) have revealed
their presence.
A rare situation that we detected it in one of the body is a bilateral relative
symmetry to a duplicated greater saphenous vein of the thigh. Individual elements
of morphology were: (a) on the right side was a shorter duplication, the anterior
duplicated trunk received the anterolateral femoral vein and the posterior
duplicated trunk received the postero-medial femoral vein, duplication continue
until the femoral vein which immediately received distal to the junction superficial
epigastric vein, superficial circumflex iliac vein and pudendal vein and in addition,
a suprapubic trunk (b) on the left side the duplication of the greater saphenous
vein is longer, continuing to the saphenofemoral junction.
Keywords: femoral vein, anterior accessory saphenous vein, saphenous femoral
junction
Kategorie: Poster
Poster 14
Rubrik: 2.Klinische Anatomie/Makroskopie
Abstract Nr.:2
Titel: Microscopic morphology concerning the duodenal intramural ganglia in
rabbits
Autoren: SISU A.(1),PETRESCU C.(1),POP E.(1),STANA L.(1),FOLESCU
R.(1),SELARU M.(1),MOTOC A.(1),RUSU M.(2),
Adressen:(1)UNIVERSITY OF MEDICINE AND PHARMACY „VICTOR
BABES”|DEPARTMENT OF ANATOMY AND
EMBRYOLOGY|TIMISOARA|ROMANIA; email:alinasisu@gmail.com;
(2)UNIVERSITY OF MEDICINE AND PHARMACY „CAROL DAVILA”|FACULTY
OF DENTISTRY|BUCHAREST|ROMANIA
Abstract:
Thuneberg (1982) distinguishes four types of interstitial Cajal cells in the visceral
wall: type I ICC - associated cells to Auerbach plexus, type II ICC - subserous
localized cells; type III ICC - circular layer cells; type IV ICC - cells in layer
external muscle. In our specimens, 12 rabbits have been dissected and then
duodenal wall has been HE stained. Were observed in the duodenal plexus
mienteric several cellular morphological types: large cells with round or oval
nucleus, hypochromic and 1-2 nucleoli, have a limited number of prolongations
and correlates morphologically with the A Park type; medium size cells with
perinuclear cytoplasmic ring; large cells with well represented cytoplasm,
hypercromic; they are few and correlates with II ICC type; cells of medium or
large size,with eccentric nuclei with abundant cytoplasm and chromatin grains;
small cells with nuclei and hypercromic embranchment, with a fine dendritic
character, star-shaped (type IV Thuneberg); cells with small nuclei and well
represented cytoplasm. In deep submucosa (corresponding to the Henle\'s
plexus) are seen in the rabbit duodenum cells: large, hypochromic nucleus with
powdery chromatin, presence of abundant cytoplasm oriented histological
diagnosis to submucous neurons rather than to the ICC; small cells which can be
rather enteric submucous interneurons. Of these cellular elements some are
emitting perivascular extensions.
Keywords: interstitial cells, neurons, submucosa;
Kategorie: Poster
Poster 15
Rubrik: 2.Klinische Anatomie/Makroskopie
Abstract Nr.:2
Titel: Mechanisms of proximal hamstring rupture in a non-athlete healthy middleaged female: a case report
Autoren: Cotofana S.(1),Tillmann B.(2),Pufe T.(3),Wambach W.(4),
Adressen:(1)Paracelsus Medical University Salzburg|Kreisklinikum Altötting,
Department of Traumasurgery, Altötting, Germany|Altötting|Germany;
email:sebastian.cotofana@pmu.ac.at; (2)Christian-AlbrechtsUniversity|Department of Anatomy|Kiel|Germany; (3)RWTH Aachen
University|Department of Anatomy and Cell Biology|Aachen|Germany; (4)|Kreisklinikum Altötting, Department of Traumasurgery|Altötting|Germany
Abstract:
This case report describes the rare case of a non-athlete healthy middle-aged
female being diagnosed with an incomplete proximal hamstring rupture. She
performed &lt;5h/week of recreational activities and never conducted competitive
sports. Regular medication or prior corticosteroid injections were negated.
Hamstring rupture was diagnosed by clinical examination and magnet resonance
imaging (MRI). Surgical refixation of the conjoint tendon of the biceps femoris and
of the semitendinosus muscle to the ischial tuberosity was performed 8 days after
the trauma. 20 weeks after surgery including directed physical rehabilitation she
experienced marginal pain and no functional impairment.
Histological analysis and immune-histochemical staining (vasculo endothelial
growth factor – receptor 2) revealed signs of fibroblast proliferation and
vasculoneogenesis indicating that repairing mechanisms and tissue remodeling
took place.
This case shows that recreational activities can lead to micro-injuries in the
proximal hamstrings without clinical signs of pain or functional impairment. These
micro-injuries induce repairing mechanisms and thus tissue remodeling leading to
consecutive tissue weakening and mechanical failure during non-adequate
trauma. Further, shows this case similar results to previous investigations made
in the filed of degenerative tendon diseases observed in Achilles tendon ruptures.
Kategorie: Poster
Poster 16
Rubrik: 2.Klinische Anatomie/Makroskopie
Abstract Nr.:2
Titel: Topographic anatomy of the infraorbital nerve, canal, and foramen
Autoren: Böhnisch M.(1),Nimtschke U.(1),Schwab W.(1),
Adressen:(1)TU Dresden|Institute of Anatomy|Dresden|Germany;
email:wolfgang.schwab@tu-dresden.de
Abstract:
The infraorbitale foramen is an important point of reference concerning dental and
surgical intervention. For blocking the infraorbitale nerve and the superior anterior
alveolar branches the anesthetic is injected directly to the opening of the
infraorbitale canal.
The aim of the present study was to describe more precisely variations in the
course of the superior anterior alveolar branches and their collateral structures in
the infraorbital canal. For the study 20 formaline-fixed sites of heads of bodydonators were examined. The documentation was done with a camera and an
electronic caliper on before-determined reference points.
In evaluating the results there could be made a classification of three main
varieties in the course of the infraorbitale nerve. The superior medial alveolar
branch was found in only 40 % of the cases. There was no infraorbitale vein
found.
The orbit was divided up into three thirds. 50 % of the foramina lied exactly at the
crossing of the first to the second third. 40 % lied within the first third. The mean
distance to the alveolar crest was 21,5 mm; the mean superior distance to the
infraorbital margin was 10,3 mm. Furthermore we found an accessory infraorbital
foramen in 30 % of the cases which was localized medial from the main foramen
and which was always smaller in size.
A more precise estimation of location and course of the dental branches allows a
better anesthesia of the nerves of the infraorbital foramen.
Kategorie: Poster
Poster 17
Rubrik: 2.Klinische Anatomie/Makroskopie
Abstract Nr.:2
Titel: Evaluation of controlled lung reperfusion strategies in a porcine model of
cardiopulmonary bypass
Autoren: Jung K.(1),Slottosch I.(2),Liakopoulos O.(2),Kuhn E.(2),Deppe
A.(2),Wahlers T.(2),Mühlfeld C.(3),
Adressen:(1)Justus-Liebieg-Universität|Anatomie und
Zellbiologie|Gießen|Deutschland; (2)Universität Köln|Herz- und
Thoraxchirurgie|Köln|Deutschland; (3)Justus-Liebig-Universität|Anatomie und
Zellbiologie|Gießen|Deutschland; email:christian.muehlfeld@anatomie.med.unigiessen.de
Abstract:
Background: Postoperative pulmonary dysfunction after cardiac surgery due to
cardiopulmonary bypass (CPB)-related ischemia/reperfusion (I/R) injury of the
lungs is a considerable source for morbidity and mortality of patients. This pilot
study tested the effects of controlled lung reperfusion strategies after CPB on
lung morphology in pigs.
Methods: During two hours of CPB the pigs were exposed to a 60-minute interval
of cardioplegic arrest with an unloaded heart and subsequent lung reperfusion.
To test the study hypothesis, the pigs were randomly allocated to one of the
following groups: group 1: uncontrolled reperfusion, group 2: controlled
reperfusion by controlling reperfusion conditions, group 3: modified controlled
reperfusion by controlling reperfusion conditions and composition of reperfusate.
The animals were observed for 4 h post-CPB and the lungs were fixed for
microscopic investigations by vascular perfusion. To evaluate the structural
preservation of the lungs, peribronchovascular and intra-alveolar edema
formation and blood-gas-barrier integrity were analyzed stereologically.
Results: No significant change could be found with regard to blood-gas-barrier,
intra-alveolar and peribronchovascular edema or any other morphological lung
parameter between the groups. Additionally functional parameters (lung
compliance, Horovitz-Index, AaDO2, intrapulmonary shunt, PVR) were
significantly deteriorated after CPB but without significant differences between
experimental groups.
Conclusion: Neither controlled, nor modified controlled reperfusion offer
significant benefit concerning injury of blood-gas-barrier as well as formation of
functionally relevant edema as compared to uncontrolled reperfusion. The
morphological findings are consistent with functional parameters demonstrating
no improvement due to controlled lung reperfusion strategies.
Kategorie: Poster
Poster 18
Rubrik: 2.Klinische Anatomie/Makroskopie
Abstract Nr.:2
Titel: Considerations regarding the importance of imaging methods in the
diagnosis of cervical neoplasm
Autoren: RADU A.(1),FRANDES C.(1),SFERDIAN M.(1),STRECU L.(1),
Adressen:(1)“VASILE GOLDIS” WESTERN UNIVERSITY OF ARAD|FACULTY
OF MEDICINE,PHARMACY AND DENTAL MEDICINE|ARAD|ROMANIA;
email:adrianadanaradu@yahoo.com
Abstract:
Introduction: The pathology of cervical malignant tumor is among the top causes
of female morbidity and mortality along with breast, lung and colorectal neoplasia,
in our coutry. Despite screening test method Babes Papanicolaou (PAP), which is
a non-invasive harvesting method, female population addressability is low, thus
that the diagnosis of cervical cancer cases is in fairly advanced stages. This
situation requires a complex course of treatment and imaging approach for future
monitoring.
Material and method: The study presents the imaging examinations performed in
patients suspected of having malignant tumors, with exo or endocervical starting
point, of which was later confirmed by clinical and pathological examination the
diagnosis of carcinoma of the cervix, in 84 patients. The patients in the study
group had performed the following imaging tests: general ultrasound and pelvic
computed tomography (CT) and magnetic resonance imaging (MRI). Radio
imaging examinations are important in tumor extensions and in highlighting the
secondary determinations.
Results and discutions: Imaging methods are an optimal way to assess the main
prognostic factors such as invasion of adjacent and regional lymph node stations
and disseminations at visceral and somatic level. These imaging methods also
allows for selection of therapeutic strategy and its moment of initiation and
implementation.
Conclusions: None of the radio-imaging methods can however determine the
type of cervical cancer, this particular aspect requiring pathological examination,
which leads us to the importance of interdisciplinary cooperation.
Key words:the cervical malignant tumors, imaging methods, the therapeutic
strategies
Kategorie: Poster
Poster 19
Rubrik: 2.Klinische Anatomie/Makroskopie
Abstract Nr.:2
Titel: Variants of brachial plexus: median and musculocutaneous nerve
Autoren: Claassen H.(1),Schmitt O.(2),Schulze M.(2),Wree A.(2),
Adressen:(1)Martin-Luther-Universität Halle-Wittenberg|Institut für Anatomie und
Zellbiologie|Halle (Saale)|Sachsen-Anhalt; email:horst.claassen@medizin.unihalle.de; (2)Universität Rostock|Institut für Anatomie|Rostock|MecklenburgVorpommern
Abstract:
Variations in the architecture of the brachial plexus are more the rule than the
exception. Among 45 arms, variations of brachial plexus were detected in 17
specimen (38%). Variants mostly concerned the fork of the median nerve (11)
and the musculocutaneous nerve (7), but seldom the ulnar nerve (1), the axillar
nerve (1) or the radial nerve (0).
In the left arm of a 79-year-old female cadaver, a fork of the median nerve was
located on profunda brachii artery which continued as a superior collateral ulnar
artery after giving origin to collateral medial and radial arteries. In the left arm of a
85-year-old female cadaver, both roots of the median nerve originated at the
lateral cord of brachial plexus. The two roots of median nerve covered the
brachial artery and joined to a fork in the distal brachial artery. Furthermore, the
ulnar nerve arose from the lateral cord of the brachial plexus and crossed axillar
artery from lateral to medial side near to the origin of subscapular artery. In a 61year-old man a branch of left musculocutaneous nerve returned to the median
nerve just after perforation of coracobrachialis muscle forming a second fork of
median nerve which covered brachial artery. At the right a similar variation of
musculocutaneous nerve was observed.
Variations of the brachial plexus can explain unexpected signs and symptoms like
paraesthesia. Due to the high frequency of plexus variations, preoperative in vivo
imaging should be interpreted more carefully before surgery of the axilla and the
upper arm.
Kategorie: Poster
Poster 20
Rubrik: 2.Klinische Anatomie/Makroskopie
Abstract Nr.:2
Titel: From MRI to anatomical specimen
Autoren: Fischer K.(1), Schwegler H.(1), Tempelmann C.(1), Voges J.(1),
Bernstein H.-G.(1), Schwab W.(2),
Adressen: (1) Otto von Guericke Uni Magdeburg|Institut für Anatomie|Klinik für
Neurologie|Klinik für Psychiatrie; email: karin.fischer@med.ovgu.de; (2)
Technische Universität Dresden|Institut für Anatomie
Abstract:
Comparative investigation of the brain employing MRI scan, macroscopic and
microscopic techniques
Interpretation of high-resolution MRI scans requires detailed knowledge of
cerebral structures. A method was developed to relate specific neuronal groups
and nuclei in vivo with macroscopic and microscopic anatomy. The head of body
donors was scanned in a 3 Tesla MRI shortly post mortem and the brain
subsequently fixed in situ. Tissue sections, corresponding to the investigated
MRI sections, were treated with BIODUR S10 as well as histologically prepared
and stained.
This Specimen allow a detailed interpretation of the MRI scan and investigation of
the artefacts during fixation and impregnation with BIODUR and paraffin.
Kategorie: Poster
Poster 21
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:3
Titel: Impact of synaptophysin on the morphology of synapses and synaptic
vesicles
Autoren: Münster-Wandowski A.(1),
Adressen:(1)Cahrité-Universitätsmedizin Berlin|Institut für Integrative
Neuroanatomie|Berlin|Deutschland; email:gudrun.ahnert@charite.de
Abstract:
Impact of synaptophysin on the morphology of synapses and synaptic vesicles
Agnieszka Münster-Wandowski, Sascha Seibert, Johannes-Friedrich Zander,
Gudrun Ahnert-Hilger
Synaptic vesicles (SV) are key organelles in neuronal communication and
characterized by a special set up of proteins. Amongst these synaptophysin is
one of the most abundant SV proteins. Despite this fact little is known about its
physiological function and synaptophysin knockout animals do not show evident
phenotypical changes. Using hippocamapal cultures from wild type and
synaptophysin knockout mice we here show that axonal and dendritic growth is
impaired in the deletion mutants. In addition synapses appear to have reduced
amounts of synaptic vesicles and SV in mutant cultures are larger than SV from
the corresponding wild type neurons. When analyzing adult hippocampal sections
by electron microscopy deletion mutants show a dramatically reduced amount of
SV especially in the mossy fiber terminals. Since synaptophysin resides on the Xchromosome only female heterozygeous exist. Milder phenotypical changes in
the morphology of mossy fibers are also observed in heterozygeous mutants,
their severity depending presumably from the inherited paternal or maternal gene.
Male-female differences are also seen when analyzing glutamate uptake into SV
prepared from mutant mice. While there are only marginal differences in the
glutamate uptake between wild types of both gender, knockout females
concentrate more glutamate compared to their male littermates. It is concluded
that synaptophysin modulates synaptic plasticity in a gender typic fashion.
Kategorie: Poster
Poster 22
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:3
Titel: Palisade endings – proprioceptors in the extraocular eye muscles?
Autoren: Lienbacher K.(1),Mustari M.(2),Messoudi A.(1),Horn A.(3),
Adressen:(1)Ludwig-Maximilians-Universität München|Institut für Anatomie und
Zellbiologie I|München|Deutschland; email:karoline.lienbacher@med.unimuenchen.de; (2)University of Washington|Washington National Primate
Research Center|Seattle|United States; (3)Ludwig-Maximilians-Universität
Müchen|Institut für Anatomie und Zellbiologie I|München|Deutschland
Abstract:
Vertebrate extraocular muscles show a highly complex anatomy, which differs in
many respects from skeletal muscles. Furthermore, there is a considerable
variation among different species with regard to the presence of proprioceptive
organs. Whereas muscle spindles and Golgi tendon organs are well developed in
sheep and pig, neither are found in cat, and only poorly developed muscle
spindles are present in human and monkey. In all vertebrates studied so far cuffs
of nerve terminals around multiply-innervated muscle fibres, termed palisade
endings (PE) are present at the myotendinous junction. While the proprioceptive
function of classical proprioceptors is well established, the function of PEs is not
clear.
PEs exhibit properties of both, motor and sensory endings. Whereas a motor
function of PEs is suggested by the expression of cholinergic markers, the vast
majority of neurotendinous junctions support a sensory function.
Recently we have shown in monkey that PEs arise from cell bodies around the
periphery of the motor nuclei of extraocular muscles together with the
motoneurons of multiply-innervated muscle fibres, which may imply a motor
function. A morphological and histochemical analysis of the peripheral neurons
around the motonuclei revealed two populations: one group of cholinergic
multipolar neurons represent the motoneurons supplying the multiple motor
innervation, one group of round calretinin-positive cholinergic neurons give rise to
PEs. If the palisade endings do have a sensory function, then their cell body
location amongst the non-twitch motoneurons would be an ideal location to
control the tension in the non-twitch extraocular muscle fibres.
Supported by DFG (HO 1639/4-3), NIH (EY013308); RR00166.
Kategorie: Poster
Poster 23
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:3
Titel: Loss of the neurotrophin receptor p75ntr alters the morphology of the
dentate gyrus
Autoren: Dokter M.(1),von Bohlen und Halbach O.(1),
Adressen:(1)Universitätsmedizin Greifswald|Institut für Anatomie und
Zellbiologie|Greifswald|Deutschland; email:oliver.vonbohlen@uni-greifswald.de
Abstract:
The p75NTR receptor binds all neurotrophins with low affinity. Depending on cosignaling with the high affinity trk-receptors, p75NTR can signal survival and
differentiation but also cell death. Since the p75NTR is highly expressed in the
dentate gyrus (DG) and since the DG is a structure capable of spinogenesis and
adult neurogenesis, we tried to identify the possible role of p75NTR in these
processes. We used mice in which the p75NTR gene was deleted. There are two
knockout mice available. In p75NTRExIII knockouts only the full-length receptor is
not expressed whereas in p75NTRExIV knockout both, the full-length and the
truncated receptor are not expressed. The use of these two knockout mice
provided the opportunity to distinguish the possible roles of the two p75NTR
receptors. Using different markers of specific stages of adult neurogenesis, we
could e.g. demonstrate an increase in the number of differentiated doublecortinpositive cells in the granule layer of the DG in p75NTR knockout mice.
Measurement of spine density in Golgi-stained sections revealed an increase in
spine density in the DG by comparing knockout and wildtype mice. Based on that,
we hypothesize that the p75NTR plays a role in neuronal differentiation within the
hippocampus.
Supported by the DFG (BO 1971/5-1) and by a grant from the Domagk-program
to M.D.
Kategorie: Poster
Poster 24
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:3
Titel: Alterations in brain architecture and dendritic spines in srgap3-deficient
mice
Autoren: Bertram J.(1),Waltereit R.(2),Bartsch D.(2),von Bohlen und Halbach
O.(1),
Adressen:(1)Universitätsmedizin Greifswald|Institut für Anatomie und
Zellbiologie|Greifswald|Deutschland; (2)Zentrum für seelische GEsundheit
(ZI)|Abt. Molekularbiologie|Mannheim|Deutschland; (2)Zentrum für seelische
Gesundheit (ZI)|Abt. Molekularbiologie|Mannheim|Deutschland;
email:oliver.vonbohlen@uni-greifswald.de
Abstract:
Some years ago, a gene, encoding for a protein called srGAP3 (or MEGAP) was
discovered, which was disrupted and functionally inactivated in a patient
displaying severe mental retardation (MR). Since srGAP3 shares homologies with
other RhoGAP-proteins, it is thought that srGAP3 plays crucial roles in neuronal
plasticity and higher brain functions. SrGAP3 -deficient mice have recently been
generated and we have started to analyze them with respect to putative
morphological alterations in the brain. The brains of srGAP3 deficient mice
display increases in total brain size and weight (increase in wet-weight of more
than 20%). This enlargement was accompanied by increases in the mean
thickness of several white mater tracts within the forebrain. Moreover, the srGAP3
deficient mice display an enlargement of the ventricles. Consistent features of
neurons in patients with MR or in several mouse models of MR are abnormal
dendritic structures and/or alterations in dendritic spine morphology. By using
computer-based reconstructions of Golgi-stained material, we found that spine
densities were not altered in hippocampal neurons (CA1 and DG) of srGAP3
deficient mice, but the individual spine length was increased. In summary, these
data support the notion that srGAP3 plays specific roles in the brain. Further
investigations will not only provide insight in the roles of srGAP3, but may also be
helpful getting insight in fundamental mechanisms involved in neuronal plasticity
and the development of mental dysfunctions.
Supported by the DFG (BO 1971/5-1)
Kategorie: Poster
Poster 25
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:3
Titel: Distribution of mas, an angiotensin-(1-7) receptor, in the murine brain
Autoren: Freund M.(1),von Bohlen und Halbach O.(1),
Adressen:(1)Universitätsmedizin Greifswald|Institut für Anatomie und
Zellbiologie|Greifswald|Deutschland; email:oliver.vonbohlen@uni-greifswald.de
Abstract:
Aside from the well-known biological active angiotensin II, other biological active
angiotensins have been discovered, including angiotensin IV and angiotensin-(17). Some years ago, it has been discovered that the Mas protooncogene encodes
an angiotensin-(1-7) receptor. Angiotensin-(1-7) is not only expressed in the
periphery but also within the brain. Based on that, we examined the distribution of
Mas within the murine brain, using an antibody directed against the 3rd
cytopasmic loop of the receptor protein. Strongest protein expression was
detected in the dentate gyrus of the hippocampus and in the piriform cortex.
However, Mas protein expression is not restricted to these areas, since Mas
immunopositive neurons were also seen in different parts of the cortex,
hippocampus, amygdala, basal ganglia, thalamus and hypothalamus. The
dentate gyrus and the piriform cortex are both capable of adult neurogenesis.
Based on the strong expression of Mas in these areas, we speculate that Mas
may interfere with adult neurogenesis. To get insight in this, we currently analyse
Mas-deficient mice. Preliminary results hinted that in Mas-deficient mice
neurogenesis is increase, in comparison to age-matched control littermates, at
least in the dentate gyrus. Thus, it may be speculated that Mas may play a role in
modulating adult neurogenesis.
Supported by the „Forschungsverbund Neurowissenschaften Greifswald“.
Kategorie: Poster
Poster 26
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:3
Titel: Characterization of affected cell populations after misexpression of thymosin
β4 in the chicken optic tectum
Autoren: Lever M.(1),Wirsching H.(2),Theiss C.(1),Brand-Saberi B.(1),
Adressen:(1)Ruhr-Universität Bochum|Institut für Anatomie und Molekulare
Embryologie|Bochum|Deutschland; email:mael.lever@googlemail.com;
(2)Universität Zürich|Labor für Molekulare Neuro-Onkologie|Zürich|Schweiz
Abstract:
Thymosin beta4 (Tbeta4) is a small, highly conserved polypeptide which is
present in nearly all mammalian cells and has multiple intra- and extracellular
functions. In addition to enabling cell migration by interacting with actin filaments,
Tbeta4 promotes cell survival and activates stem cells e.g. in damaged tissues
and by that enhances wound healing and tissue repair. However, until now, the
role of Tbeta4 in neurogenesis / gliogenesis has not been precisely described.
In the present study, we first analyzed the physiological Tbeta4 expression in the
embryonic optic tectum of the chicken and then electroporated plasmids for overexpression or knockdown of Tbeta4. In contrast to the neural stem cells of the
ventricular zone, which did not show a positive Tbeta4 signal, early neurons in the
tectal plate, and, at later developmental stages, all neuronal layers revealed a
prominent Tbeta4 expression. Knockdown of Tbeta4 inhibited the growth of the
transfected hemispheres, whereas Tbeta4 over-expression led to a remarkable
cell proliferation, causing the formation of folds and groves resembling gyri and
sulci which are specific of higher vertebrates and are normally absent in chicken.
After characterizing the expression of specific immunohistochemical markers for
the radial glial (Vimentin), neuronal (DCX, Tuj-1) and glial (GFAP) cell population
throughout the normal development of the chicken optic tectum using confocal
laser scanning microscopy, we compared these results to the expression of these
markers in the electroporated optic tectum.
Our findings account for a major step towards understanding the important role of
Tbeta4 in glio- and neurogenesis.
Kategorie: Poster
Poster 27
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:3
Titel: Time-lapse images of cytoskeletal rearrangement in neuronal growth cones
Autoren: Olbrich L.(1),Brand-Saberi B.(1),Theiss C.(1),
Adressen:(1)Ruhr-Universität Bochum|Institut für Anatomie und Molekulare
Embryologie|Bochum|Deutschland; email:laura.olbrich@gmail.com
Abstract:
Time-lapse images of cytoskeletal rearrangement in neuronal growth cones
Laura Olbrich, Beate Brand-Saberi, Carsten Theiss
Institut für Anatomie & Molekulare Embryologie, Ruhr-Universität Bochum,
Universitätstr.150, 44780 Bochum, Deutschland
VEGF is a dimer polypeptide originally known in the context of angiogenesis as a
growth factor for vascular endothelial cells. In the neuronal system VEGF has a
positive impact on the survival of neuronal stem cells, astrocytes and microglia,
and is additionally is known to prevent growth cone collapse.
The aim of the present study is to investigate the effect of VEGF on the neuronal
growth cone, a highly specialized structure at the tip of growing axons of neurons.
This very motile structure was first described in the 19th century and translates
guidance signals into cytoskeletal arrangements to route the axon to its final
destination. Morphologically, the growth cone is divided into peripheral (P),
central (C) and transition (T) regions, each characterized by specific cytoskeletal
proteins.
With the aid of confocal laser scanning microscopy we were able to detect the
cytoskeletal rearrangements subsequent to stimulation with VEGF. Therefore we
used chicken-DRG neurons to study growth cone dynamics without and after
exposure to VEGF in vitro. To illustrate the protein allocation, we used primary
neurons following microinjection with a combination of GFP-neurofilament and
RFP-actin. We investigated alterations in shape, outgrowth and motility,
comparing the time-lapse images under various conditions. Additionally, we
repeated our studies using progesterone, another effector in the nervous system.
To corroborate these findings, we evaluated the outgrowth of treated and
untreated organotypic DRG-cultures using immunohistochemistry and compared
shape and width of neuronal growth cones under various conditions.
Kategorie: Poster
Poster 28
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:3
Titel: Cortical plasticity under the influence of progesterone in developing purkinje
cells
Autoren: Wessel L.(1),Meller K.(2),Brand-Saberi B.(1),Theiss C.(3),
Adressen:(1)Ruhr-Universität Bochum|Institut für Anatomie und Molekulare
Embryologie|Bochum|Deutschland; email:lisa-wessel@web.de; (2)RuhrUniversität Bochum|Abteilung für Cytologie|Bochum|Deutschland; (3)RuhrUniversität Bochum|IInstitut für Anatomie &amp; Molekulare
Embryologie|Bochum|Deutschland
Abstract:
New findings over the past decade have shown that different cells in the brain are
able to synthesise steroid hormones de novo from cholesterol, called
neurosteroids.
Recently, Purkinje cells (PC), the principal neurons of the cerebellar cortex, have
been identified as a major site for neurosteroid production in vertebrates. In
addition to pregnenolone and pregnenolone sulfate, progesterone is synthesized
in developing PCs. But in contrast to these neurosteroids, actively synthesised
progesterone is only detectable during the neonatal period, when the integration
of the cerebellar cortex increases dramatically. Thus, progesterone is thought to
be involved in the formation of the cerebellar neuronal circuit by promoting
dendritic growth, spinogenesis and synaptogenesis in developing PCs.
It is likely that progesterone has an age-related diverging effect on the cortical
plasticity of the cerebellar circuit. To test this hypothesis we compared different
parameters such as dendritic length or spine density at given time points of the
cerebellar development before and after stimulation with progesterone. According
to the physiological course of the progesterone concentration in PC, our
preliminary results show that treatment with progesterone achieves the highest
effects on spine density and dendritic length during the early stages of
development.
A further working point of our study was to investigate the outcome of
progesterone antagonism with mifepriston for the developing cerebellar PC.
Referring to this interrogation we treated cerebellar rollertube slices at given time
points with mifepriston and compared well-defined parameters of treated and
untreated slice cultures.
Kategorie: Poster
Poster 29
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:3
Titel: Vegf and its influence on cultured astrocytes
Autoren: Wuestefeld R.(1),Meller K.(2),Brand-Saberi B.(2),Theiss C.(1),
Adressen:(1)Ruhr-Universität Bodhum|Institut für Anatomie und Molekulare
Embryologie|Bochum|Deutschland; email:ricky.wuestefeld@t-online.de; (2)RuhrUniversität Bochum|Institut für Anatomie &amp; Molekulare
Embryologie|Bochum|Deutschland
Abstract:
The purpose of the present study was to investigate the effects of Vascular
Endothelial Growth Factor (VEGF) on cell dynamics in primary astrocytes, gap
junctional intercellular communication (GJIC) and cell proliferation. VEGF is
known as a dimeric polypeptide which potentially binds to two receptors, VEGFR1 and VEGFR-2, however, many effects of VEGF are mediated by VEGFR-2, e.g.
forced cell migration, angiogenesis, actin polymerization, and cell proliferation.
Recently, it has been shown that in case of hypoxia, injury and ischemia VEGF is
upregulated to stimulate cell proliferation in endothelial cells and angiogenesis.
Besides this, VEGF reveals a potent therapeutical target for averting tumor
vascularization, emerging in bevacizumab, the first humanized anti-VEGF-A
antibody for treating recurrent Glioblastoma multiforme.
In order to expand and combine our knowledge about VEGF effects in glial cells,
we cultivated rat astrocytes in medium containing VEGF for several days. To
investigate the effects of VEGF on GJIC, we microinjected neurobiotin into a
single cell and monitored dye-spreading into adjacent cells. These experiments
showed that VEGF significantly enhances astrocytic GJIC compared to controls.
Cell proliferation measured by BrdU-labelling also revealed a significant increase
of astrocytic mitotic rates subsequent to VEGF exposure. To study cell-dynamics
of astrocytes with relation to VEGF treatment, we additionally transfected
astrocytes with LifeAct-RFP. Live-cell imaging and quantitative analysis of these
cells with aid of confocal laser scanning microscopy revealed higher process
motility of VEGF treated astrocytes. In conclusion, VEGF strongly affects GJIC,
cell proliferation, and process motility in cultured astrocytes.
Kategorie: Poster
Poster 30
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:3
Titel: Diurnal sorting of vglut1 between vesicular and plasma membrane
compartments: is the vglut1/endophilin interaction the key?
Autoren: Richter K.(1),Ahnert-Hilger G.(2),
Adressen:(1)Charité Berlin,|Zentrum für Anatomie. Institut für Integrative
Neuroanatomie|Berlin|Deutschland; email:karin.richter@charite.de; (2)Charité
Berlin|Zentrum für Anatomie, Institut für Integrative
Neuroanatomie|Berlin|Deutschland
Abstract:
Three structurally related vesicular transporters (VGLUT1 -3) are responsible for
glutamate loading of synaptic vesicles (SV). Studies using VGLUT1 or VGLUT2
deletion mutants revealed the copy number per vesicle being crucial for synaptic
efficiency (Weston et al., 2011)
When analysing SV prepared at different times of the day diurnal changes of the
VGLUT amounts are observed (Yelamanchili et al., 2006). These variations are
probably due to a diurnal switch of VGLUTs between the vesicular and the
plasma membrane (Darna et al., 2009).
Recently, it has been shown, that VGlut1 but not VGlut2 interacts with the SH3
domain of Endophilin, a protein, involved in clathrin-dependent endocytosis
(Voglmaier et al., 2006).
In order to investigate whether an oscillating binding of VGlut1 to Endophilin
forms the basis of the circadian oscillation in the amount of vesicular VGlut1,
brain extracts of mice entrained in a
12h light/12h dark-cycle or 12h dark/12h dark-cycle were analysed. Pull down
assays using GST fusion protein of the SH3 domain of Endophilin, showed an
oscillating binding pattern of VGlut1 (in ratio to Dynamin) to Endophilin. Deletion
of the period gene Period 2 necessary for clock resetting in Per2BRDM1 mice
resulted in the loss the varying interactions between VGlut1/Dynamin and
Endophilin. The variations in the VGLUT1/endophilin interactions in wild type
mice however do not mirror the oscillations of the amounts of VGLUT1 on SV
suggesting other and/or additional proteins being involved in the unique endocytic
retrieval of VGLUT1.
Kategorie: Poster
Poster 31
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:3
Titel:Compartmentalization of prominin-1/cd133 within the brain of mammalian
and non-mammalian vertebrate species
Autoren: Jászai J.(1),Corbeil D.(1),
Adressen:(1)TU Dresden|Tissue Engineering|Dresden|Deutschland;
email:jozsef.jaszai@biotec.tu-dresden.de
Abstract:
In mammals, neuroepithelial progenitors and those of later stages with radial glial
features in the fetal/postnatal brain as well as adult neural stem cells can be
prospectively isolated based on the cell surface expression of prominin-1
(CD133). In contrast, the cellular and molecular characterization of neural
progenitors in non-mammalian vertebrates exhibiting significant constitutive
neurogenesis and inherent self-repair ability is particularly hampered by the lack
of suitable cell surface markers. Genes encoding for prominin-1 are potentially
present in all metazoan genomes, and we have recently identified them from the
major non-mammalian vertebrate model organisms including zebrafish, axolotl
and chick (Jászai et al. 2011, PLoS One 6:e17590). It is currently unknown,
however, whether these molecules show any degree of conservation as for their
association with mammalian neurogenic niches within central nervous system
(CNS). Moreover, the phenotypic properties of activated neural progenitors during
provoked neurogenesis in the regenerating CNS are poorly understood. By
mapping the spatiotemporal expression of prominin-1–related transcripts by nonradioactive in situ hybridization combined with PCNA or BrdU-labeling of actively
dividing neural progenitor cells within the intact brain, we uncovered a markedly
conserved distribution of this molecule among vertebrate species. Furthermore,
we observed that cells in the regenerating spinal cord of injured axolotl were
characterized by enhanced expression of prominin-1. Collectively, our novel
information gained about molecular properties of neural progenitors within the
intact and injured CNS might enable us to answer open questions regarding to
the significant inherent regenerative capacity of the non-mammalian CNS.
The study was supported by the Deutsche Forschungsgemeinschaft (TRR83 No.
6; SFB655 B3 and CO298/5-1).
Kategorie: Poster
Poster 32
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:3
Titel: Atomoxetine alters leves of synaptic proteins in adult and fetal rat brain
Autoren: Udvardi P.(1),Liebau S.(1),Dreyhaupt J.(2),Fegert J.(3),Böckers
T.(1),Ludolph A.(3),
Adressen:(1)Universität Ulm|Institut für Anatomie und
Zellbiologie|Ulm|Deutschland, Baden-Württemberg; (2)Universität Ulm|Institut für
Epidemiologie und Medizinische Biometrie|Ulm|Deutschland, BadenWürttemberg; (3)Universität Ulm|Klinik für Kinder- und
Jugendpsychiatrie/Psychotherapie|Ulm|Deutschland, Baden-Württemberg
Abstract:
The pathogenesis of psychiatric disorders as well as the mechanisms of the
pharmacological treatments are still not completely understood. Attentiondeficit/hyperactivity disorder (ADHD) is the most frequently diagnosed
neuropsychiatric disorder in childhood. As atomoxetine is the first non-stimulant
compound licensed for the treatment of ADHD, we addressed the issue of its
impact on the developing brain in rodents. Here we demonstrate that acute
atomoxetine treatment (3 mg/kg, i.p.) of pregnant Sprague Dawley rats (E12 E18) significantly shifted expression levels of genes coding for synaptic proteins
solely in the dams brains, whereas synaptic protein levels were altered in both
adults and fetuses. In the dams brain atomoxetine reduced levels of
monoaminergic transporters in prefrontal cortex and striatum (STR). In the adult
STR we furthermore identified significantly increased levels of mRNA and protein
of the vesicular glutamate transporter 1, the presynaptic vesicle marker
synaptophysin (Syp) and the ADHD candidate gene Snap25. Syp levels were
also increased in the mesencephalon (MES) of adults. However, in the fetal brain,
atomoxetine elicited significantly reduced protein levels of N-methly-D-aspartate
receptor subunits (NR1, NR2B) and increased histone 2B protein levels in both
MES and STR. In summary these data clearly demonstrate that atomoxetine’s
cellbiological effects are not limited to the well-characterized inhibition of
norepinephrine reuptake, but also imply development-dependent action on brain
biochemistry. Furthermore our in vivo study strongly indicates, that atomoxetine’s
impact may also alter neuronal and synaptic plasticity.
Kategorie: Poster
Poster 33
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:3
Titel: Association of the motor protein kif26b with the postsynaptic density protein
abi-1
Autoren: Heinrich J.(1),Proepper C.(1),Boeckers T.(1),
Adressen:(1)Ulm|Anatomie und Zellbiologie|Ulm|Germany;
email:jutta.heinrich@uni-ulm.de
Abstract:
Morphogenesis and several aspects of a functional cell are dependent on
regulated intracellular transport mechanisms, which are mostly based upon
cytoskeletal components and motor proteins such as kinesins that actively
transport membranous organells and protein complexes along microtubules in an
ATP-dependent manner.
Over the last years we characterised the postsynaptic density (PSD) protein
Abelson interacting protein 1 (Abi-1) which is important for dendrite branching,
spine morphology and synapse formation and a direct interaction partner of the
master scaffolding protein ProSAP2/SHANK3. Interestingly, we could show that
Abi-1 translocates from the PSD to the nucleus upon NMDA receptor stimulation.
To better understand the molecular mechanisms behind these transport
processes, we performed a yeast two-hybrid screen of a fetal human brain library
using a complete cDNA of Abi-1 as bait. Among several candidate genes we
found a partial cDNA of Kif26b, an N-kinesin that walks along microtubules. In
several experiments we were able to show that Kif26b is localised in neurons of
the hippocampus as well as Abi-1 and that both proteins specifically interact with
each other. Further experiments are set up to characterize Kif26b in neurons and
to elucidate the role it may play in synapto-nuclear transport processes.
Kategorie: Poster
Poster 34
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:3
Titel: Morphological and gene expression changes of NSC-34 motor neuron-like
cells during differentiation and estrogen treatment.
Autoren: Maier O.(1),Brück S.(1),Beyer C.(1),Johann S.(1),
Adressen:(1)RWTH Aachen|Neuroanatomie|Aachen|Deutschland;
email:oliver.maier@rwth-aachen.de
Abstract:
Morphological and gene expression changes of NSC-34 motor neuron-like cells
during differentiation and estrogen treatment.
Oliver Maier, Stefan Brück, Cordian Beyer, Sonja Johann.
Abstract
Aims: NSC-34 are a murine neuroblastoma X spinal cord hybrid cell line and
represent an in vitro model for spinal cord motor neurons. This subclass of
neurons are highly affected in motoneuron diseases like Amyotrophic Lateral
Sclerosis (ALS). 17beta-oestradiol is well known by its action on the cholinergic
system where it can influence the expression patterns of several cholinergic
enzymes. Therefore, differentiated NSC-34 cells were treated with 17betaestrogen and specific estrogen receptor agonists to characterise the intracellular
pathway of estrogen action on the neuronal cholinergic system.
Methods: NSC-34 cells were differentiated using retinoic treatment. Differentiation
was analysed by gene expression of neuronal differentiation markers and by
counting of neurites. Effects of 17beta-oestradiol on cholinergic proteins in
differentiated NSC-34 was measured by real-time PCR and Western Blotting.
Morphological changes were analysed by immunocytochemistry and
immunofluorescence.
Key findings: The expression of cholinergic and neurofilament proteins was highly
upregulated in the cells during the differentiation process and increased neurite
outgrowth and synaptic formations were observed. Furthermore, 17betaoestradiol treatment of differentiated NSC-34 cells significantly increased the
expression of several cholinergic enzymes, including Choline Acetyltransferase.
Kategorie: Poster
Poster 35
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:3
Titel: Indirect effects of ciliary neurotrophic factor (cntf) produced in olfactory
ensheathing cells (oec) on olfactory sensory neuron (osn) neurite growth in
cocultures
Autoren: Bömmel H.(1),Kneitz S.(2),Asan E.(1),
Adressen:(1)Universität Würzburg|Institut für Anatomie und
Zellbiologie|Würzburg|Deutschland; (2)Universität Würzburg|IZKF Microarray
Core Unit|Würzburg|Deutschland; email:esther.asan@mail.uni-wuerzburg.de
Abstract:
OEC, glial cells of the olfactory nerve, promote and guide axonal outgrowth.
CNTF, a neurotrophic factor implicated in neuroprotection and axonal
regeneration, supposedly participates in mediating this effect. Surprisingly, OSN
displayed significantly increased neurite length if cultured on CNTF-deficient
compared to wildtype OEC, and application of CNTF neutralizing antibodies to
the culture medium enhanced neurite length in rat OSN/OEC and CN/OEC
cocultures while addition of recombinant CNTF to CNTF-deficient cocultures
decreased neurite length, suggesting that CNTF mediated the effect.
In OSN monocultures, the growth substrate (laminin or merosin) significantly
influenced neurite growth while adding CNTF at concentrations sufficient to
reduce neurite length in cocultures was without any effect. To study whether
differences in extracellular matrix (ECM) composition could be responsible for
differential neurite growth in OSN/OEC cocultures we cultured OSN from both
genotypes on extracellular matrix (ECM) derived from wildtype and CNTFdeficient OEC monocultures. Mean neurite length was significantly increased on
ECM from CNTF-deficient OEC. Microarray analysis revealed differences in the
expression of numerous genes, including genes coding for ECM components, in
OEC cultures from different genotypes. This finding could be confirmed for
selected genes by RT-PCR using mRNA from cultured OEC and olfactory bulbs
of wildtype and CNTF-deficient animals. Additionally, application of CNTF
neutralizing antibodies to wildtype OEC cultures induced analogous changes of
mRNA expression.
Our data suggest that CNTF from OEC exerts an indirect effect on OSN neurite
outgrowth in OEC cocultures, possibly via modulating production of specific axon
growth-related molecules in OEC.
Kategorie: Poster
Poster 36
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:3
Titel: Alphapix (arhgef6) inhibits caleb/ngc-mediated dendritic tree and spine
complexity
Autoren: Schumacher S.(1),Johannes S.(1),Rosenberger G.(2),Kutsche K.(2),
Adressen:(1)Universitaet Ulm|Institut fuer Molekulare und Zellulaere
Anatomie|Ulm|Deutschland; email:stefan.schumacher@uni-ulm.de;
(2)Universitaetsklinikum Hamburg-Eppendorf|Institut fuer
Humangenetik|Hamburg|Deutschland
Abstract:
Patients with mental retardation-linked disorders often display abnormalities in
dendritic tree and spine architecture. Several genes which encode for proteins
functionally connected to cytoskeletal rearrangements have been found to be
mutated in patients suffering from mental retardation. One of these proteins is
alphaPIX (ARHGEF6), a member of the guanine nucleotide exchange factors
(GEFs) that mediates activation of Rho GTPases. Previously we showed that
alphaPIX interacts with CALEB/NGC, a neural member of the EGF family which
induces dendritic tree and spine complexity in neurons. We demonstrated that
knockdown of alphaPIX potentiates CALEB/NGC-stimulated dendritic branching.
We now present data which are concordant with a model of alphaPIX function, in
which alphaPIX acts upstream of CALEB/NGC to inhibit CALEB/NGC-mediated
dendritic tree and spine complexity. We propose that the interaction between
alphaPIX and CALEB/NGC could be of functional relevance for the shaping of
dendritic trees and spines in mental retardation-linked disorders.
Supported by the Deutsche Forschungsgemeinschaft (DFG, SCHU 1406/3-1).
Kategorie: Poster
Poster 37
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:3
Titel: Neocortical development in Bcl11a/Ctip1 mutant mice
Autoren: Wiegreffe C.(1),Nelles E.(1),Heiland C.(1),Glatz S.(1),Jenkins
N.(2),Copeland N.(2),Britsch S.(1),
Adressen:(1)Ulm University|Institute of Molecular and Cellular
Anatomy|Ulm|Germany; email:christoph.wiegreffe@uni-ulm.de;
(2)Proteos|Institute of Molecular and Cell Biology|Singapore|Republic of
Singapore
Abstract:
In the neocortex, the zinc-finger protein Bcl11a/Ctip1 is broadly expressed by
cells within the cortical plate from E12.5 onwards and can be found in all
neocortical layers during later stages of development. Using forebrain-restricted
ablation of Bcl11a we found this factor to play important roles in neocortex
development. Postnatally, the cortex of Bcl11a mutant mice is significantly
reduced in size and shows a disorganization of cortical layers suggesting roles of
this molecule during migration and differentiation of cortical neurons. To better
understand the molecular mechanisms behind the observed phenotype, we
performed comparative transcriptome analyses of mutant and control brains.
Candidate target genes are further analyzed on a molecular (in situ hybridization,
RT-qPCR) and biochemical level (ChIP). To functionally evaluate the target
genes for their effect on neuronal migration/differentiation we have established a
technique that combines ex utero electroporation with organotypic slice culture.
This powerful in vitro technique is easy to use, generates highly reproducible
results, allows for direct observation of migrating neurons, and efficient screening
of comparatively large numbers of genes for their effect on cortical neuronal
migration/differentiation. We will generate knockdown and overexpression
constructs that contain a GFP reporter to perform loss-of-function, gain-offunction, and rescue experiments in Bcl11a mutants. With our approach we plan
to uncover the molecular mechanism of Bcl11a function in projection neurons
during cortical development and discover new genes involved in this process.
Kategorie: Poster
Poster 38
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:3
Titel: Endocytic trafficking of fibroblast growth factor receptor 1 in human glioma
cells
Autoren: Irschick R.(1),Karp G.(1),Klimaschewski L.(1),
Adressen:(1)Medizinische Universität Innsbruck|Division für
Neuroanatomie|Innsbruck|Österreich; email:Regina.Irschick@i-med.ac.at
Abstract:
Fibroblast growth factor receptor 1 (FGFR1) is an important receptor tyrosine
kinase responsible for glioblastoma growth. Binding of the ligand FGF triggers the
intracellular Ras/Raf/Erk pathway leading to cell proliferation. Down-regulation of
the receptors and inhibition of their signaling pathways represents a new
treatment strategy for glioblastoma patients. We are investigating the endocytic
trafficking of FGFR1 in the human glioma cell line U373. It is now clear that
receptor endocytosis alone is not sufficient for attenuation of the signaling
cascade, but fusion of the endosomes with multi-vesicular-bodies and
subsequent trafficking to lysosomes is necessary for receptor degradation.
The glioma cells were transfected with fluorescent FGFR1 constructs, and
colocalization of the receptor was determined with vesicular markers for early,
late and recycling endosomes as well as lysosomes. Fluorescence microscopy
was performed with a confocal microscope (Leica SP5) and with structured
illumination techniques (Zeiss AxioObserver ApoTome), and 3D image analysis
was performed with Huygens Deconvolution Software.
Our findings suggest that FGF-2 enhances colocalization of FGFR1 with early,
late and recycling endosomes and lysosomes. Treatment with the lysosomal
inhibitor leupeptin results in receptor accumulation in late endosomes and
lysosomes. Analysis of vesicle distributions shows an accumulation of recycling
endosomes in the perinuclear region, while late endosomes and FGFR1containing vesicles are homogeneously distributed throughout the cell.
Taken together, analysis of cellular FGFR1 distribution in glioma cells under
different treatment conditions leads to a better understanding of receptor
trafficking and could help to develop new strategies to reduce FGFR1-mediated
tumor growth.
Kategorie: Poster
Poster 39
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:3
Titel: Genetic analysis of the zinc finger transcription factor bcl11b/ctip2 in adult
neurogenesis and maturation of hippocampal neurons
Autoren: Fischer J.(1),Schwegler H.(2),Jenkins N.(3),Copeland N.(3),Simon
R.(1),Britsch S.(1),
Adressen:(1)Ulm|Molecular and Cellular Anatomy|Ulm|Germany; (2)Otto-vonGuericke39129|Anatomy|Magdeburg|Germany; (3)Singapore|Molecular and
Cellular Biology|Singapore|Singapore
Abstract:
The hippocampus, in particular the dentate gyrus, is one of only two brain regions
where adult neurogenesis occurs. With advancing age, the proliferative activity of
hippocampal stem cells and their neuronal differentiation capacities continuously
decline, resulting in a dramatic, reduction in neurogenesis and the ability to learn
new memory tasks. We have previously shown that Bcl11b/CTIP2, a Krueppellike zinc finger transcription factor, is specifically expressed in postmitotic granule
cell neurons of the dentate gyrus throughout life. Using conditional gene targeting
in mice we demonstrated that Bcl11b is essential for development of the dentate
gyrus. Forebrain-specific ablation of Bcl11b results in a hypoplastic dentate gyrus
caused by reduced progenitor cell proliferation and arrested differentiation of
postmitotic granule cell neurons. As a consequence, Bcl11b mutants exhibit
severe deficits in learning behavior. To specifically determine functions of Bcl11b
in the adult and aging dentate gyrus we generated an inducible mouse line using
the Tet-off system under the control of the CaMKIIa promoter. Here we present
data analyzing the proliferation and differentiation of hippocampal neurons as well
as the learning behavior of animals exhibiting an induced ablation of Bcl11b in
dentate granule cells only in adult age.
Kategorie: Poster
Poster 40
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:3
Titel: Bcl11b function unexpectedly involves action of a common cell adhesion
molecule
Autoren: Venkataramanappa S.(1),Schwegler H.(2),Jenkins N.(3),Copeland
N.(3),Simon R.(1),Britsch S.(1),
Adressen:(1)Ulm|Molecular and Cellular Anatomy|Ulm|Germany; (2)Otto-vonGuericke|Anatomy|Magdeburg|Germany; (3)Singapore|Molecular and Cellular
Biology|Singapore|Singapore; email:ruth.simon@uni-ulm.de
Abstract:
The hippocampus has an important function in learning, memory storage and
spatial navigation. The dentate gyrus of the hippocampus is also one of only two
brain centers with continuing neurogenesis in adulthood. The development of the
dentate gyrus is characterized by specific phases involving specific molecular
pathways.
Bcl11b/CTIP2, a Krueppel-like zinc finger transcription factor is expressed in
postmitotic granule cells of the dentate gyrus early on during development
throughout adulthood. Previously our lab demonstrated an important function of
Bcl11b in the regulation of progenitor cell proliferation as well as neuronal
differentiation. Loss of Bcl11b expression results in a hypoplastic dentate gyrus
caused by reduced progenitor cell proliferation as well as an arrest of neuronal
differentiation leading to impaired learning and memory behavior. Transcriptome
analysis identified several candidate genes, which were verified by RT-PCR, in
situ hybridization as well as Chromatin-immunoprecipitation. A common cell
adhesion molecule was identified as a potential direct Bcl11b target gene.
Analyzing mutants for this gene revealed an overlapping phenotype when
compared to Bcl11b mutants suggesting a direct function in the development of
the dentate gyrus. Our data link a transcriptional program to a molecule involved
in cell-cell adhesion as well as cytoskeleton structures.
Kategorie: Poster
Poster 41
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:3
Titel: Regulation of small heat shock proteins via micrornas in hippocampal
neurons?
Autoren: Bartelt-Kirbach B.(1),Golenhofen N.(1),
Adressen:(1)Universität Ulm|Institut für Anatomie und
Zellbiologie|Ulm|Deutschland; email:britta.bartelt@uni-ulm.de
Abstract:
Non-physiogical conditions trigger the stress response of cells, leading to the
induction of heat shock proteins. The family of small heat shock proteins (sHsps)
are molecular chaperones preventing irreversible aggregation of partially unfolded
proteins. We could show previously that four of the eleven familiy members are
upregulated in rat hippocampal neurons under various stress conditions,
indicating a possible neuroprotective role. Interestingly, HspB5 (alpha-Bcrystallin) showed an upregulation on protein level but not on mRNA level. This
prompted us to investigate the possible involvement of microRNAs in the
regulation of this sHsp. MicroRNAs are small non-coding RNAs which bind to the
3’-UTR of their target mRNAs, leading to a translational inhibition. Cultured rat
hippocampal neurons were subjected to heat or simulated hypoxic stress and the
expression of 388 rat microRNAs compared to matched controls by array
hybridization. 22 microRNAs were downregulated more than two-fold after heat
stress while 58 microRNAs showed downregulation after simulated hypoxia.
Seven of these microRNAs were common to both treatment groups. In contrast,
29 microRNAs are upregulated more than two-fold after heat stress, 77 after
hypoxia with an overlap of 6 microRNAs in both groups.
In conclusion, microRNAs show a distinct reaction to stress conditions, hinting at
a strong involvement of this class of molecules in the cellular stress response. Of
particular interest are the microRNAs which show a common downregulation after
heat and hypoxic stress because these are the ones which might upregulate
specific heat shock proteins, leading finally to increased cellular survival at stress
conditions.
Kategorie: Poster
Poster 42
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:3
Titel: Conditional knock-out of tgfbr2 in telencephalon of mice causes defects in
neurovascular development
Autoren: Büttner N.(1),Wahane S.(1),Vogel T.(1),
Adressen:(1)Albert-Ludwigs-Universität Freiburg|Institut für Anatomie und
Zellbiologie, Abteilung Molekulare Embryologie|Freiburg|Deutschland;
email:nicole.buettner@anat.uni-freiburg.de
Abstract:
Transforming growth factor beta (Tgf-beta) has an important role in embryonic
and adult organisms. To investigate functions of Tgf-beta in forebrain
development in vivo, we used a FoxG1-cre mouseline to conditionally knock-out
Tgfbr2. FoxG1 is expressed in progenitors and neurons of the telencephalon.
FoxG1cre/+; Tgfbr2flox/flox mice display severe haemorrhages mostly in the
telencephalon and diencephalon beginning around E13.5. The embryos die
between E16 to E17, when the entire forebrain is infiltrated with blood. These
massive bleedings are not only due to the knock-out of Tgfbr2, but also depend
upon reduced expression of FoxG1. This is supported by the observations that
other conditional knock-out mutants of Tgfbr2 in telencephalon did not show a
neurovascular phenotype in our previous studies.
To further investigate the neurovascular defect we characterise these mutants
with regard to angiogenesis, neurogenesis, proliferation and apoptosis. Blood
vessels show an atypical appearance especially in the subcortical area. In this
area we also observe fewer vessels and less branching. Cell junctions and basal
membrane seem to be properly developed. As FoxG1 is not expressed in the
endothelial cells itself, Tgfbr2 is not knocked-out in these cells. This indicates that
most likely signals coming from neurons or progenitors mediate this vascular
defect. We are currently analysing molecular signals in this context with neuronal
origin that affect endothelial development and maturation. We hypothesize that
excess Tgf-beta signalling through loss of feedback inhibition results in
endothelial mesenchymal transition that is accompanied by leaky and less
branched vessels.
Kategorie: Poster
Poster 43
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:3
Titel: Deficiency for p75ntr enhances early as well as late phase of ltp in the
amygdala
Autoren: Baldus M.(1),von Bohlen und Halbach O.(1),
Adressen:(1)Universitätsmedizin Greifswald|Institut für Anatomie und
Zellbiologie|Greifswald|Deutschland; email:oliver.vonbohlen@uni-greifswald.de
Abstract:
The neurotrophin receptor P75NTR is thought to have an inhibitory influence on
neurite growth, cell differentiation and survival of basal forebrain cholinergic
neurons (BFCNs), which in turn modulate synaptic transmission of brain
structures involved in learning and memory.
P75NTR deficient mice display increased numbers of cholinergic neurons and we
could show that p75NTR deficient mice have a higher cholinergic innervation of
the hippocampus and amygdala as compared to control littermates. Concerning
the hippocampus, deletion of p75NTR induces increases in spine densities and
dendritic branching patterns in area CA1; however, enhanced as well as
unchanged long-term potentiation (LTP) in area CA1 has been reported for
p75NTR deficient mice.
Since deletion of p75NTR has an impact upon the innervation of the amygdala,
we examined the effects of a deletion of p75NTR upon LTP in the amygdala. We
found that both, early- as well as late-LTP, are significantly enhanced in the
lateral nucleus of the amygdala of p75NTR-deficient mice. Whether these effects
are attributed to the altered cholinergic innervation or to alterations in the
neurotrophin signalling in the p75NTR deficient mice remain to be clarified.
Supported by the DFG (BO 1971/5-1)
Kategorie: Poster
Poster 44
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:3
Titel: Interrelations between monoaminergic afferents and interneurons in the
mouse laterobasal amygdala: investigations in wildtype and serotonin
transporter (5-htt)-deficient mice
Autoren: Renninger C.(1),Schwert H.(1),Bonn M.(1),Schmitt A.(2),Asan E.(1),
Adressen:(1)Universität Würzburg|Institut für Anatomie &amp;
Zellbiologie|Würzburg|Deutschland; email:christoph.renninger@uniwuerzburg.de; (2)Universität Würzburg|Klinik für Psychiatrie, Psychosomatik und
Psychotherapie|Würzburg|Deutschland
Abstract:
The amygdala is implicated in processing and memory of emotional stimuli.
Monoaminergic afferents and intrinsic inhibitory interneuronal systems modulate
information processing in the main target area of sensory inputs to the amygdala,
the laterobasal nuclei. Previous studies using immunohistochemistry and in situ
hybridization (ISH) in rats documented that laterobasal NPY-producing
interneurons are densely and moderately innervated by serotonergic and
dopaminergic afferents, respectively, and express different subtypes of serotonin
receptors. The aim of the present work was to adapt methods established in the
rat for application on mouse brain tissue to enable analyses of the systems in
mice with targeted genetic modifications of molecules involved in monoaminergic
transmission.
Immunohistochemistry showed similar distribution of NPY-immunoreactive(ir)
neurons in the laterobasal amygdala in rats and mice as well as an analogously
dense innervation of these interneurons by serotonergic and tyrosine
hydroxylase-ir, presumably dopaminergic fibers. The same was true for
parvalbumin-ir interneurons. Additionally, ISH for serotonin (5-HT) receptor
subtypes 1A, 2C and 3A mRNA was successfully applied on sections of mouse
amygdala and documented the presence of these receptor subtypes in the
laterobasal nuclei also of this species. Qualitatively, interneuron distribution and
dopaminergic innervation of the laterobasal amygdala were similar for wildtype
and 5-Htt-deficient mice. Further analyses of the monoaminergic innervation and
comparative quantitative studies in genetic mouse models for emotional
dysregulation in humans are now possible and will provide a basis for
interpretation of the functions of monoaminergic and interneuronal amygdaloid
systems and their interrelations in brain emotion circuits.
Kategorie: Poster
Poster 45
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:3
Titel: The role of neuregulin 1 (nrg 1) in the hippocampal morphology and motor
and learning capabilities
Autoren: Kräusel S.(1),Schwegler H.(1),Garratt A.(2),Birchmeier C.(2),Roskoden
T.(1),
Adressen:(1)Otto-von-Guericke-Univ.|Institut für
Anatomie|Magdeburg|Deutschland; (2)Max-Delbrück-Centrum für Molekulare
Medizin (MDC)|Developmental Biology / Signal Transduction|BerlinBuch|Deutschland
Abstract:
Neuregulin1 (NRG1) is a member of a protein family consisting of 4
transmembrane proteins with an EGF-like signaling domain. NRG1 plays an
important role in different organs. In the CNS NRG1 plays a role in neuronal
migration, axonal pathfinding, synaptic function and also affects oligodendrocytes.
Recently, NRG1 has been shown to be involved in the pathogenesis of
schizophrenia. Some patients show white matter abnormalities and, additionally,
a high expression of NRG1 was found in brains of people suffering from
schizophrenia.
In the presented study we investigated different types of behavior (Open field,
spatial learning in Radial maze, Elevated plus maze and Beam walking for motor
skills), between heterozygous NRG1-knockout (KO) and their wild type (WT)
littermates. Furthermore, we studied hippocampal glutamatergic and cholinergic
markers using Timm- and AChE-staining.
In the radial maze, both genotypes showed significant learning performance but
no between-genotype differences. Neither in the open field nor in the elevated
plus maze significant differences between groups were found. NRG1
heterozygous mice showed a small though not significant motor deficit indicated
by more sliding in beam walking.
The morphological analysis resulted in significantly higher density of intra- and
infrapyramidal mossy fibers in the CA3-region of KO. No differences were found
for AChE fiber-density. These results demonstrate NRG1 affects the
glutamatergic transmission but not the cholinergic in the hippocampus. Small
differences in beam walking might indicate involving of NRG1 in signal processing
in nervous system. The results exhibit morphological but no behavioral
differences induced by a NRG1 deficit in mice.
Kategorie: Poster
Poster 46
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:3
Titel: Effects of prg 1 on olfaction – behavioral and morphological differences
Autoren: Schneider P.(1),Schwegler H.(1),Nitsch R.(2),Vogt J.(2),Roskoden T.(1),
Adressen:(1)Otto-von-Guericke-Univ.|Institut für
Anatomie|Magdeburg|Deutschland; (2)Johannes Gutenberg Univ.|Instituts für
Mikroskopische Anatomie und Neurobiologie|Mainz|Deutschland;
email:thomas.roskoden@med.ovgu.de
Abstract:
Plasticity related gene1 (PRG1) is an integral membrane protein located in
postsynaptic glutamatergic synapses. It controls signaling pathways of
phosphorylated lipid substrates. A lack of PRG1 leads to a lower ability to analyze
sensory information and in juvenile mice to epileptic discharge.
To analyze the effects of PRG1 on sensory pathways we used homozygous
PRG1-knockout (KO) and their wild type littermates (WT). Since olfaction is the
most important sensory system in mice, we exposed the animals to a predator
odorant 2,5-dihydro-2,4,5-trimethylthiazoline (TMT) in a small box (45x45cm) and
observed the behavioral reactions elicited by TMT. After TMT-exposure we
analyzed c-fos activation in brain areas associated with olfaction using
immunohistochemistry.
The deficient PRG1KO shows significantly lower freezing activity and an
enhanced jumping activity than WT animals. This indicates that KO shows the
fear reaction when exposed to the odor but the sensory processing of odor is less
efficient than in WT.
The morphological analysis of the olfactory pathway revealed an enhanced
number of c-fos reactive cells in PRG1KO compared to WT after TMT-exposure.
These differences were most pronounced in the olfactory tubercle, mediodorsal
thalamus and olfactory bulb. The olfactory tubercle is involved in the unconscious
part of the olfactory pathway which is involved in freezing behavior. The lack of
PRG1 results in a higher amount of phosphorylated lipids postsynaptically and
modulates excitation. This leads to an overexcitability of glutamatergic neurons
and might explain the differences in behavior after TMT-exposure and the
observed stereotypic phenomenons.
Kategorie: Poster
Poster 47
Rubrik: 3.Neuroanatomie/Neurobiologie
Abstract Nr.:3
Titel: Attenuation of autophagy by a knockdown of beclin-1 enhances the
sensitivity of hippocampal neurons to amino acid starvation and induced an aifdependent apoptosis
Autoren: Kim M.(1),Rami A.(1),
Adressen:(1)Wolfgang Goethe-University|Dr. Senckenbergische Anatomie,
Institute of Cellular and Molecular Anatomy|Frankfurt|Germany;
email:rami@em.uni-frankfurt.de
Abstract:
Beclin 1 is involved in the regulation of autophagy in mammalian cells and in the
facilitation of autophagic cell death by regulated binding to the prototypic
apoptosis inhibitor Bcl-2. In addition, Beclin 1 is monoallelically deleted in many
forms of sporadic breast, ovarian and prostate cancer. Mice that carry
heterozygous disruption of Beclin 1 have a high incidence of spontaneous
tumours, and cells with reduced Beclin 1 expression exhibit reduced autophagic
activity.
This study examined the potential role of Beclin-1 in an autophagic response in
hippocampal HT22 neurons challenged with amino acid starvation (AAS). AAS in
wild type cultures induced light chain-3 (LC-3)-immunopositive and
monodansylcadaverine (MDC) fluorescent dye-labelled autophagosome
formation. In addition, AAS induced neuronal death without affecting caspase-3-,
AIF- or HtrA2-levels. In contrast, in Beclin-1 knockdown HT22 neurons, AAS
induced: 1) a dramatic upregulation of AIF, 2) a caspase-independent neuronal
death 3), a decrease in the LC3-II/LC3-I ratio, 4) and reduced accumulation of
autophagosomes.
Collectively, this study shows that the autophagic machinery is inducible in
cultured hippocampal HT22 neurons subjected to AAS. Our data further show
that inhibition of autophagy by a knockdown of Beclin-1, enhanced susceptibility
to proapoptotic signals induced by AAS und underlines that autophagy is per se a
protective than a deleterious mechanism.
This study was supported by a grant of the “Adolf-Messer-Stiftung” to A.R.
Kategorie: Poster
Poster 48
Rubrik: 4.Zellbiologie
Abstract Nr.:
Titel: Contribution of vascular wall-resident progenitor cells (VW-EPCs) to tumor
vascularization
Autoren: Mertins S. ,Kleff V. ,Hohn H-P, Singer B.B., Klein D., Ergün S.
Adressen: University of Duisburg-Essen|Institut for Anatomie|Essen|Germany
Abstract:
Purpose: To evaluate the contribution of the VW-EPCs to tumor vascularization
and the disintegration of the vasculogenic zone (VZ).
Methods: We examined human urothelial tumors of different staging using
immunohistochemical staining for CD34 followed by statistical analysis in order to
judge the pattern of CD34+ cells in the VZ of the blood vessels and evaluated the
number of blood vessels with and without intact VZ with respect to their distance
to tumor tissue. We also used a mouse model to evaluate the migration and
differentiation of VW-EPCs during tumor vascularization and to distinguish VWEPCs from BM-derived EPC. C57Bl/6 mice were lethally irradiated and
intravenously transplanted with 2x106 murine EGFP-expressing BM cells from
EGFP-5Nagy donor mice. After hematopoietic reconstitution, tumor cells (B16F10
cells) were subcutaneously transplanted into the flank of the mice. Tumor and
surrounding tissues were removed and analyzed.
Results: The degree of disintegration varied in dependence to the distance
between blood vessels and tumor tissue. Within the tumor tissue all blood vessels
were found to be without VZ and blood vessels in close vicinity to tumor tissue
exhibited a partially disintegrated VZ almost all large and middle sized blood
vessels far from the tumor tissue displayed an intact VZ. Comparable results
were obtained for glioblastoma, sarcoma and testicular tumors. In experimental
tumors, blood vessels were found to contain both endothelial cells only positive
for CD34 and those only positive for CD31 while only few endothelial cells of
tumor vessels expressed both CD31 and CD34.
Conclusions: Our results suggest that VW-EPCs are apparently involved directly
in new vessel formation and may serve as a local source for tumor
vascularization. Thus, the VW-EPCs have to be considered in future strategies for
anti-angiogenic tumor therapy.
Kategorie: Poster
Poster 49
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: Decellularization of porcine achilles tendon and recellularization using
human hamstring tendon-derived tenocytes
Autoren: Lohan A.(1),Stoll C.(1),Denner A.(1),Albrecht M.(1),Ertel W.(1),John
T.(1),Schulze-Tanzil G.(1),
Adressen:(1)Charité Berlin|Klinik für Unfall- und
Wiederherstellungschirurgie|Berlin|Deutschland; email:anke.lohan@charite.de
Abstract:
Regenerative approaches by using natural decellularized xenogenic extracellular
matrix (xECM) for tissue reconstruction attract increasing interest. The aim of this
study was to test whether porcine Achilles tendon can be decellularized and
subsequently this cell-free xECM could be recellularized in vitro using human
hamstring tendon-derived tenocytes.
Decellularization of porcine Achilles tendons and the effect on the xECM was
controlled
using
histological
stainings
(hämatoxylin
eosin
[HE],
diamidinophenylindole [DAPI]) before reseeding with primary human hamstring
tenocytes for 1-2 weeks using statical and dynamical seeding techniques.
Subsequently, vitality testing (fluorescein diacetate/propidium iodide staining) and
histological stainings (HE, alcian blue, resorcin fuchsin, DAPI) document the
success of recellularization. Further, species differences between human and
porcine tendons were characterized.
Decellularization was successfully performed using an adapted protocol, but lead
to some loss of proteoglycans and ECM structure. Porcine Achilles tendon could
be recellularized with human tenocytes. However, cell distribution remained
mostly inhomogeneously, with cell accumulations at the margin of the constructs.
The vitality of the immigrating tenocytes decreased after one week. Porcine and
human tendons differed in regard to morphology, cellularity, ECM structure and
proteoglycan content, whereby porcine tendons were characterized by a higher
cell, proteoglycan and elastic fiber content.
The present study depictures some key steps and problems of optimizing porcine
tendon xECM de- and recellularization using human tenocytes to prepare a cellbased scaffold for tendon repair.
Kategorie: Poster
Poster 50
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: Expression and regulation of cxcl12 and its receptors in rat c6 glioma cells
Autoren: Hattermann K.(1),Mrugalla K.(2),Mentlein R.(1),Held-Feindt J.(2),
Adressen:(1)Christian-Albrechts-Universität zu Kiel|Anatomisches
Institut|Kiel|Deutschland; email:k.hattermann@anat.uni-kiel.de; (2)ChristianAlbrechts-Universität zu Kiel|Klinik für Neurochirurgie UKSH Campus
Kiel|Kiel|Deutschland
Abstract:
The chemokine CXCL12 / stromal cell-derived factor-1 (SDF-1) is produced by
fibroblasts in various organs, and regulates cell migration processes in health and
disease. In human gliomas, the most malignant brain tumor, CXCL12 mediates
migration and proliferation of distinct tumor cell subpopulations via its renown
receptor CXCR4. Recently, a further receptor for CXCL12 was de-orphanized and
termed CXCR7. This novel receptor for CXCL12 is elevated in several human
tumor types, and amongst these in gliomas. Here, we analyze the expression,
regulation and function of CXCL12 and its both receptors in the well characterized
rat glioma cell line C6. CXCL12 and its receptor CXCR7 were detected at notable
levels by quantitative RT-PCR, while CXCR4 was low. In response to low
concentrations of the alkylating agent temozolomide which is commonly used in
glioma therapy, CXCL12 and its receptors were significantly upregulated. To
show functional relevance of CXCR7, we first analyzed surface and intracellular
expression of CXCR7 by immunocytochemistry followed by functional assays
where CXCL12 reduced anti-proliferative and pro-apoptotic effects of
temozolomide via CXCR7. These data indicate that CXCL12 and its receptors,
especially CXCR7, may contribute to chemotherapy resistance in glioma cells.
Kategorie: Poster
Poster 51
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: Cytokinesis failure causes multipolar mitosis in a new cell line established
from human glioblastoma
Autoren: Telentschak S.(1),Bloch W.(2),Addicks K.(1),Klinz F.(1),
Adressen:(1)Köln|Anatomie I|Köln|Germany; email:stelents@smail.uni-koeln.de;
(2)Deutsche Sporthochschule Köln|Institut für Kreislaufforschung und
Sportmedizin, Abt. Molekulare und Zelluläre Sportmedizin|Köln|Germany
Abstract:
Glioblastoma multiforme (GBM) is the most frequent as well as malignant human
brain tumour and still associated with a very poor prognosis.
Immunofluorescence experiments have demonstrated that multipolar spindles
exist in cultured GBM cells, however no studies were performed to analyze the
fate of daughter cells born through multipolar mitosis. Using time lapse video
microscopy we have analysed mitosis in a new cell line established from GBM
and found that at early passage number about 3.4% of cells underwent multipolar
mitosis, of which 1.8% were tripolar, 0.7% tetrapolar and 0.8% of higher or
undefinable polarity. Many daughter cells born by multipolar mitosis were viable
and performed subsequent rounds of bipolar mitosis, whereas other did not enter
mitosis again or failed to complete cytokinesis and attempted later to initiate a
new multipolar mitosis. Pedigree analysis of mitotic events revealed that in many
cases a bipolar mitosis with failed cytokinesis occurs prior to a multipolar mitosis.
This mechanism is the predominant route to generate multinucleated giant cells.
In summary, cells of our newly established glioblastoma cell line performed
rounds of polyploidization and subsequent attempts of depolyploidization as well
as centrosomal amplification and subsequent efforts of deamplification. Amplified
centrosomes and polyploidy may lead to merotelic chromosome attachments in
bipolar and multipolar mitosis of multinucleated giant cells and provide the basis
for chromosomal instability resulting in loss of heterozygosity, chromosomal
lagging and loss or gain of chromosomes.
Our results indicate that multipolar mitosis may be an important mechanism to
generate heterogeneity of glioblastoma cells.
Kategorie: Poster
Poster 52
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: Characterization and differentiation potential of equine adipose tissuederived mesenchymal stem cells
Autoren: Raabe O.(1),Shell K.(1),Reich C.(1),Würtz A.(1),Damm A.(1),Crispens
C.(1),Wenisch S.(1),Arnhold S.(1),
Adressen:(1)Justus-Liebig Universität Giessen|Institut für Veterinär-Anatomie, Histologie und -Embryologie|Giessen|Deutschland;
email:oksana.raabe@vetmed.uni-giessen.de
Abstract:
Adipose tissue-derived stem cells (ADSCs) are the adult stem cells with have
reached a high level of scientific attention during the recent years. They represent
a subpopulation of adult stem cells that can be successfully used for tissue
engineering in veterinary medicine, especially in degenerations of the
musculoskeletal system in horses.
In this study we investigated the morphology, population doubling time (PDT),
differentiation potential and molecular biological properties of the equine ADSCs
from subcutaneous fat tissue.
Expression of Oct4, Nanog, CD90, CD105 was assessed by RT-PCR.
Differentiation potential was analyzed by histological stains and RT-PCR
analyses.
ADSCs showed a good proliferative potential and can differentiate into the
adipogenic and osteogenic lineage. Expression of peroxisome proliferator
activated receptor (Pparγ2) mRNA was positive in adipogenic lineages and
alkaline phosphatase (AP), phosphoprotein (SPP1), Runx2 and osteocalcin (OC)
mRNA were positive in osteogenic lineages.
These results demonstrate a multiple differentiation potential of equine ADSCs
and their high value for tissue engineering applications in equine veterinary
medicine.
Kategorie: Poster
Poster 53
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: Establishing an efficient protocol for the generation of donor specific ips
cells
Autoren: Linta L.(1),Stockmann M.(1),Kleger A.(2),Böckers T.(1),Liebau S.(1),
Adressen:(1)Uni Ulm|Institut für Anatomie und Zellbiologie|Ulm|Deutschland;
email:leonhard.linta@uni-ulm.de; (2)Uni Ulm|Klinik für Innere Medizin
I|Ulm|Deutschland
Abstract:
Human induced pluripotent stem cells (iPS cells) are considered to be a valuable
tool for studying in vitro embryonic development, cell differentiation and tissue
formation. Additionally, they provide the possibility to study various pathogenetic
pathways and may be used for approaches in regenerative medicine. In this
context, somatic cells are reprogrammed by forced expression of several factors,
known as the Yamanaka factors. The arising iPS cells share all abilities of
embryonic stem cells, such as self renewal, differentiation into cells of all three
germ layers and the ability of teratoma formation.
We focused our work on establishing a noninvasive protocol to obtain adult
somatic cells and reprogram them into iPS cells with a high efficiency leading to
an improved success rate of iPS generation. Therefore we chose keratinocytes
from plucked human hair as a starting population and reprogrammed them by
lentiviral transduction of an excisable construct containing all reprogramming
factors as a single transcript. Subsequently, infected keratinocytes are transferred
to a feeder cell layer of fibroblasts that are known to support the viability and
metabolism of the reprogramming cells. When testing various culture conditions
and feeder cell types during the reprogramming we found that the use of rat
embryonic fibroblasts (REF) is superior to the commonly used mouse embryonic
fibroblasts (MEF) in the reprogramming process leading to a significant increase
of iPS colonies, displaying all hallmarks of pluripotency. Together, the presented
method of human iPS cell generation, the utilized cell source and the newly found
feeder cell type proved to have a very high success rate and was used to
generate numerous lines from healthy and diseased donors.
Kategorie: Poster
Poster 54
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: Glycohistochemical characterization of an immortalized human sebaceous
gland cell line (sz95)
Autoren: Rodler D.(1),Schneider M.(2),Habermann F.(1),Zouboulis
C.(3),Sinowatz F.(1),
Adressen:(1)Universitaet Muenchen|Department of Veterinary
Sciences|Muenchen|Deutschland; email:d.rodler@anat.vetmed.unimuenchen.de; (2)Universitaet Muenchen|Gene Center|Muenchen|Deutschland;
(3)The Free University of Berlin|Department of Dermatology|Berlin|Deutschland
Abstract:
Immortalized human sebocytes of the SZ95 cell line offer unique possibilities for
investigations on the physiology of the sebaceous gland and its role in skin
diseases. Moreover, this model can be used to evaluate the biologic activity of
different compounds on sebaceaous gland cells in vitro. It has been shown in
previous studies that SZ95 cells synthesize highly glycosylated molecules like
epithelial sialomucin (MAM-6) and milk fat globulin-2. In the present study a
glycohistochemical characterization of SZ95 cells has been performed using a
panel of 13 different FITC-labeled lectins, which bind to specific sugar structures.
The glycohistochemical staining pattern before and after retinoic acid induced
differentiation of the SZ95 has been evaluated. We could demonstrate a distinct
staining of the cells with WGA, succinylated WGA, PHA-E, LCA, PNA and PSA.
Although the phenotype of the cells changed during the 7 days of induced
differentiation, no significant alterations of the lectin binding was observed.
Kategorie: Poster
Poster 55
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: Organization of CD44 and the microfilament system in cells of the
macrophage lineage during osteoclast differentiation
Autoren: Hund R.(1),Glenske K.(1),Tryankowski E.(1),Schwartz-Albiez
R.(2),Wenisch S.(1),
Adressen:(1)Justus-Liebig-Universität Gießen|Institut für Veterinär-Anatomie, Histologie und -Embryologie|Gießen|Deutschland;
email:roman.hund@vetmed.uni-giessen.de; (2)Deutsches
Krebsforschungszentrum|Tumor Immunology Program|Heidelberg|Deutschland
Abstract:
CD44 is a membrane protein, plays a role in fusion, and might be associated with
a special actin subdomain, called the podosomal core. Little information is
available about the spatiotemporal pattern of CD44 synthesis regarding
coinciding changes of the actin cytoskeleton which occur during differentiation
and fusion of mononucleated macrophages into polykaryons. In order to
investigate the dynamics of CD44 synthesis and expression we have performed
appropriate cell cultures experiments.
The human peripheral blood mononuclear cells were obtained by density gradient
and cultivated in medium containing M-CSF and RANKL. Cells were fixed at days
1, 7 and 10.
CD44 was detected by immunofluorescence (CD44 antibody mouse anti human,
AbDserotec). Phalloidin-fluorescence-isotiocyanat (Sigma-Aldrich) was used to
visualize the actin cytoskeleton.
The results revealed perinuclear CD44 signalling and Phalloidin staining in
macrophages at the beginning of culturing. In the course of culturing the
perinuclear CD44 signals became stronger and could be also detected along the
plasma membranes of the cells which formed clusters in order to fuse. When
polykaryons were formed punctate phalloidin staining pointed towards the
formation of podosomes. At the end of culturing, actin belts were formed along
the plasma membranes which still revealed strong CD44 staining.
The results demonstrate that differentiation of macrophages into polykaryons in
vitro is characterized by typical spatiotemporal distribution patterns of CD44 and
actin filaments. The results should help to understand the mechanisms underlying
the foreign body reaction which occurs in vivo after implantation of any medical
device.
This work is supported by the DFG/Collaborative Research Center Transregio 79.
Kategorie: Poster
Poster 56
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: Role of desmoglein 2 for keratinocyte adhesion
Autoren: Hartlieb E.(1),Spindler V.(1),Waschke J.(1),
Adressen:(1)Ludwig-Maximilians University Munich|Institute of Anatomy and Cell
Biology, Department I|München|Deutschland; email:jens.waschke@med.unimuenchen.de
Abstract:
Desmosomes provide intercellular adhesion required for integrity of epithelial and
a variety of non-epithelial tissues. In stratified epithelia such as the epidermis, the
transmembrane cadherin family proteins desmogleins (Dsg) 1-4 and desmocollins
(Dsc) 1-3 build the core of desmosomes by interacting in the intercellular space
and by being tethered to intracellular plaque proteins. Although in keratinocytes
several isoforms of desmosomal cadherins are co-expressed, the contribution of
the specific isoforms to overall cell cohesion is unclear. Here, we characterized
the role of Dsg2 in comparison to Dsg3, the latter of which is known to be
essential for keratinocyte adhesion. In the HaCaT keratinocytes both Dsg2 and
Dsg3 are expressed in confluent cells. Incubation of keratinocytes with a
monoclonal antibody (AK23) targeting the Dsg3 adhesive domain led to profound
loss of cell adhesion as detected by dispase-based dissociation assays. In
contrast, a Dsg2 antibody which is also known to directly interfere with Dsg2
trans-adhesion had no effect. To substantiate these differences we performed
siRNA-mediated knockdown of Dsg2. Again, cell-cell adhesion was slightly
reduced. Protein levels of other desmosomal cadherins were not significantly
altered by reduced Dsg2 expression except of Dsg3 which was slightly
diminished. Interestingly, subsequent incubation with AK23 led to drastically
enhanced keratinocyte cell dissociation in Dsg2 knockdown cells compared to
cells transfected with non-targeting siRNA. These experiments indicate that
specific desmosomal cadherins contribute differently to keratinocyte cohesion.
Since Dsg2 may compensate at least partially for loss of Dsg3 function, an
additive mechanism of desmosomal cadherins for cell adhesion is likely.
Kategorie: Poster
Poster 57
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: Assessment of in vitro biocompatibility of orthodontic mini screws in human
gingival fibroblasts and saos-2 cell cultures
Autoren: Finke H.(1),Fischer-Brandies H.(2),Es-Souni M.(2),
Adressen:(1)Christian-Albrechts-Universität Kiel|Anatomisches
Institut|Kiel|Deutschland; email:h.finke@anat.uni-kiel.de; (2)Universitätsklinikum
Schleswig-Holstein, Campus Kiel|Klinik für Kieferorthopädie|Kiel|Deutschland
Abstract:
Background and aims:
Mini screws were developed during the last decades to provide a skeletal
anchorage for orthodontic treatments. Although there are various clinical surveys,
the reasons of failure have not been revealed completely. The aim of this study
was to analyse differences in surface structure and alloy and their effects on the
in vitro biocompatibility.
Material and methods:
Experiments were performed with four different mini screws of comparable size:
tomas-pin (Dentaurum), OrthoEasy pin (Forestadent), Dual Top (Jeil
Medical/Promedia) and LOMAS screw (Mondeal). A scanning electron
microscope (SEM) with energy dispersive X-ray spectroscopy (EDX) was used to
detect differences between the four products concerning their surface structure
and the alloy composition. For the assessment of cytotoxicity MTT assays and
agar overlay assays with neutral red dye were performed.
Results:
The SEM and EDX data showed slight surface modifications in some screws.
There was a slightly reduced viability in the MTT assays after contact with tomaspin compared to the LOMAS screw. The tomas-pin, OrthoEasy pin and Dual Top
led to slight decolourations of osteoblast like cells in the agar overlay assays.
Discussion:
The results presume differences in the biocompatibility of tested mini screws. Due
to the small degree of difference to negative controls none of the products should
be classified as cytotoxic. For a clear conclusion concerning advantages of one
product over another, further clinical and in vitro tests are required.
Kategorie: Poster
Poster 58
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel:A model to study repair processes in the murine tracheal epithelium in vivo
Autoren: Döring K.(1),Knebel G.(1),König P.(1),
Adressen:(1)Universität zu Lübeck|Institut für Anatomie|Lübeck|Deutschland;
email:doering@anat.uni-luebeck.de
Abstract:
The naphthalene model of damage and repair of the airway epithelium is well
established but thought to be restricted to small intrapulmonary airways. We
aimed to determine if naphthalene can also be used to study repair processes in
murine tracheal epithelium. Explanted mouse tracheae were kept in HepesRinger solution at 37°C and were incubated with naphthalene (200 µg/ml) for 2 h.
In vivo, a dose of 300 mg/kg naphthalene was injected intraperitoneally. Tracheae
were explanted and fixed at 12 h, 24 h, 48 h, 4 d as well as 15 d and examined
using semi thin sections, scanning and transmission electron microscopy and
immunohistochemistry. Ex vivo application of naphthalene resulted in detachment
of secretory cells from the basement membrane leaving islets of ciliated cells. In
vivo after 12 h, the detachment of few secretory as well as ciliated cells was
observed. After 24 h, the epithelium was destroyed with the majority of secretory
and ciliated cells being lost. After 48 h, no ciliated and secretory cells were found.
However, basal cells still covered the basement membrane. 4 d after application
of naphthalene, a continuous epithelium was detected and short cilia were
observed. After 15 d, repair of the epithelium was completed and cilia reached
their initial length. In conclusion, naphthalene also damages large airways and
results in rapid repair but in contrast to small airways, not only secretory but also
ciliated cells are lost. Thus, the naphthalene model is also suitable to study repair
processes of the murine tracheal epithelium.
Kategorie: Poster
Poster 59
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: The chick eye as a new model for the limbal stem cell niche
Autoren: Chankiewtz V.(1),Chankiewitz E.(2),Eulitz M.(3),Theiß C.(3),Meller
D.(2),Steuhl K.(2),Brand-Saberi B.(3),
Adressen:(1)Ruhr- Universität Bochum|Institut Anatomie und Molekulare
Embryologie|Bochum|Deutschland; email:verena.chankiewitz@rub.de;
(2)Universität Duisburg-Essen|Zentrum für Augenheilkunde/ Universitätsklinikum
Essen|Essen|Deutschland; (3)Ruhr-Universität Bochum|Institut für Anatomie und
Molekulare Embryologie|Bochum|Deutschland
Abstract:
The maintenance of a transparent ocular surface requires the continuous
regeneration of the corneal epithelium. For this a special stem cell population is
located at the corneal limbus. The limbal stem cells form a small subpopulation of
about 1% of the basal cells in the limbal epithelium. Limbal stem cell insufficiency
will impair proper visual function. Therefore the reconstruction of the ocular
surface by replacement with ex vivo expanded stem cells is an important clinical
approach. The efforts to develop advanced substitutes by tissue engineering are
limited because of an incomplete molecular characterization of the limbal stem
cells and their niche. One reason for this is the lack of an adequate animal model.
The aim of or study was to establish the chick eye as a new model for the limbal
stem cell niche.
We found high similarities between chick and human anatomy in cornea and
limbal region. Especially the Bowman’s layer can be clearly distinguished in chick
histological sections in contrast to mammalian models. Transmission electron
microscopic studies visualized a small number of basal located cells which could
be identified as putative stem cells. The corresponding area investigated by
scanning microscopy comprises structures similar to limbal crypts. Specific
antibodies against several stem cell markers detected a spatial and temporal
dynamic cellular pattern during the formation of the limbal stem cell niche.
From or findings we conclude that the chicken eye will be an easy to handle,
exact definable model to investigate limbal stem cell biology.
Kategorie: Poster
Poster 60
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: Co-culture of mesenchymal stem cells with primary tendon-derived
tenocytes promotes tenogenic differentiation in vitro
Autoren: Schneider P.(1),Buhrmann c.(2),Mobasheri A.(3),Busch F.(4),Matis
U.(5),Shakibaei M.(4),
Adressen:(1)LMU-München|Institue of antomy|München|D; (2)LMUMünchen|Institute for anatomy|München|D; (3)University of Nottingham|Faculty of
Medicine and Health Sciences|Nottingham|UK; (4)LMU-München|Institute of
anatomy|München|D; (5)LMU-München|Clinic of Veterinary Surgery|München|D;
email:mehdi.shakibaei@med.uni-muenchen.de
Abstract:
Objective:
Mesenchymal stem cells (MSCs) have potential applications in regenerative
medicine, because they are pluripotent and have the capacity to differentiate into
many different cell types. However, the capacity of MSCs for differentiating into
tenocytes in vitro and has not been investigated. The aim of this study was to
investigate the tenogenic potential of MSCs when cultured with primary tenocytes
in different ratios.
Materials and methods:
Co-cultures of tenocytes and MSCs were mixed in different ratios in monolayer
and in high-density cultures. MSCs were also cultured with spent media from
primary tenocytes.
Results:
Tenogenesis was induced in high-density co-cultures of tenocytes and MSCs in
different ratios and through cultivation with the spent media from primary
tenocytes. Ultrastructural evaluation of high-density co-cultures using electron
microscopy demonstrated tenogenesis. Immunoblotting confirmed expression of
collagen type I/III, decorin, tenomodulin, betta1-Integrin, MAPKinase pathway
(Shc, Erk1/2) and scleraxis in the co-cultures. In monolayer co-cultures, MSCs
and tenocytes actively searched for cell–cell contact leading to cell proliferation
and actively exchanged vesicles, which were labelled by using
immunfluorescence and immunogold techniques, suggesting the uptake and
interchange of soluble factors produced by the MSCs and/or tenocytes.
Conclusion:
MSCs represent an attractive cell source for tendon repair strategies. This study
suggests that MSCs may be considered as a cell source for tendon regenerative
medicine and therapeutic approaches.
Kategorie: Poster
Poster 61
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: Behaviour and differentiation potential under the influence of 3% and 21%
oxygen tension of equine adipose tissue derived stem cells
Autoren: Shell K.(1),Raabe O.(1),Wenisch S.(1),Arnhold S.(1),
Adressen:(1)Justus-Liebig Universität Giessen|Institut für Veterinär - Anatomie, Histologie und -Embryologie|Giessen|Deutschland;
email:Katja.N.Faquet@vetmed.uni-giessen.de
Abstract:
Equine multipotent mesenchymal stem cells (MSCs) can be isolated from
different tissues and are capable to differentiate into various organ progenitor
cells. Physiological oxygen conditions in diverse tissues in vivo are hypoxic, even
so this fact is often overlooked in standard culture conditions, where the oxygen
tension is usually normoxic. Here, equine adipose derived MSCs were used to
analyze their behaviour and differentiation potential into the adipogenic,
osteogenic and chondrogenic lineage under 3 per cent and 21 per cent -oxygen
tension.
Hypoxia-inducible-factor-1alpha is an indicator for hypoxic stress sensed by cells.
Its expression was similar under both oxygen conditions, which is probably a sign
for low oxygen tension being sensed as normoxic by those stem cells.
Adipogenesis and chondrogenesis showed better results under 3 per cent
oxygen; for osteogenesis an oxygen tension of 21 per cent was more effective.
This knowledge may help to improve conditions and results of stem cell
differentiation and consequently their application in tissue engineering.
Kategorie: Poster
Poster 62
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: Cellular and subcellular localization of pten in odontoblasts
Autoren: Korkmaz Y.(1),Klinz F.(2),Beikler T.(1),Bloch W.(3),Raab W.(1),Addicks
K.(2),
Adressen:(1)Heinrich-Heine-University|Department of Operative Dentistry,
Periodontics and Endodontics|Düsseldorf|Germany; email:yueksel.korkmaz@uniduesseldorf.de; (2)University of Cologne|Department I of
Anatomy|Cologne|Germany; (3)German Sport University|Department of
Molecular and Cellular Sport Medicine|Cologne|Germany
Abstract:
Knowledge about the signaling processes involved in the odontoblasts
differentiation is important to the understanding of dentin repair in the case of
caries. Phosphatase and tensin homologue (PTEN) is a dual lipid-protein
phosphatase that dephosphorylates phosphatidylinositol 3,4,5-triphosphate to
phosphatidylinositol 3,4-biphosphate and inhibits phosphatidylinositol 3-kinase
(PI3K)-Akt/PKB-dependent cell proliferation and cell differentiation. Deletion of
PTEN in knockout mice only in osteoblasts induces an increase in formation of
bone indicating an important role of PTEN and PI3K-Akt/PKB signaling in bone. In
our earlier report, we have described phosphorylation of Akt/PKB in odontoblasts.
In order to clarify the cellular and subcellular localization of PTEN in odontoblasts,
decalcified frozen-sections of rat jaws and human molars were incubated with
mouse monoclonal and rabbit polyclonal antibodies against PTEN. In comparison
to the moderate staining of PTEN in rat molar odontoblasts, higher staining
intensities of PTEN were detected in odontoblasts of the rat incisivus. In
odontoblasts of the human molars, PTEN was detected with a moderate staining
intensity in cytoplasm but with a higher staining intensity in the nucleus. These
data indicate that PTEN is localized in odontoblasts and gives evidence for an
involvement of Akt/PKB-dependent odontoblast differentiation. Currently we are
studying the effects of Akt/PKB phosphorylation in dentin formation by deletion of
PTEN only in odontoblasts using Cre-mediated recombination.
Kategorie: Poster
Poster 63
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: Induction of osteoporosis in the rat model - characterization of bone and
bone marrow derived mesenchymal stem cells
Autoren: Goergen J.(1),Hempel U.(2),Raabe O.(1),Heiß C.(3),Wenisch
S.(1),Arnhold S.(1),
Adressen:(1)Justus- Liebig- Universität Gießen|Institut für Veterinär-Anatomie, Histologie, und -Embryologie|Gießen|Deutschland;
email:julia.goergen@vetmed.uni-giessen.de; (2)TU Dresden, Medizinische
Fakultät, Medizinisch-Theoretisches Zentrum|Institut für Physiologische
Chemie|Dresden|Deutschland; (3)Universitätsklinikum Giessen und Marburg
GmbH|Klinik und Poliklinik für Unfallchirurgie|Gießen|Deutschland
Abstract:
We compare three different ways of induction of osteoporosis in rats overlooking
the bone metabolism and the characteristics of bone marrow derived stem cells.
One method of induction is ovariectomy and application of steroids as a model for
steroid- induced osteoporosis. Ovariectomy and a calcium deficient diet serve as
a model for the postmenopausal osteoporosis. A sham-operated group serves as
a control. Samples of these groups are taken after one, three and twelve months
respectively. In order to reconstruct the senile osteoporosis samples are taken
after eighteen months. Characterization of the bone metabolism is performed by
analyzing the gene expression of Rank, RankL, OPG and M-CSF. Mesenchymal
stem cells are isolated from the bone marrow and characterized regarding
morphology and capacity for proliferation and differentiation. Furthermore woundhealing-assays are carried out to study the cell migration in vitro.
Differences between the groups could be observed regarding to the cell
morphology. Particularly cells harvested after three months show a higher
number of the broad and flattened cell morphology. This cell type is described as
the phenotype of aged stem cells which are in a state of replicative senescence
(Tuli R et al, 2003).Oil-Red-O staining showed a higher number of fat vacuoles in
cells from animals that received the calcium deficient diet. These findings could
indicate a cell differentiation rather towards the adipogenic lineage.
This work is supported by the DFG/Collaborative Research Center Transregio 79.
Tuli R et al., Characterization of multipotent progenitor cells derived from human
trabecular bone, Stem Cells 2003; 21:681-693
Kategorie: Poster
Poster 64
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: Alpha 10-subunit of nAChR and nicotine-induced cilia-driven particle
transport in mouse respiratory epithelium
Autoren: Weitz A.(1),Hartmann P.(1),Faulhammer P.(1),Boseva I.(1),Alvarez
T.(1),Ibanez-Tallon I.(2),Kummer W.(1),Krasteva G.(1),
Adressen:(1)Justus-Liebig-Universität|Institut für Anatomie und
Zellbiologie|Giessen|Deutschland; email:Ariane.Weitz@anatomie.med.unigiessen.de; (2)-|Max-Delbrück-Centrum für Molekulare
Medizin|Berlin|Deutschland
Abstract:
Mucociliary clearance, the cleaning mechanism of the airways, relies on ciliadriven particle transport. It is driven by an intrinsic cholinergic system including
muscarinic receptor M3. The role of nicotinic receptors (nAChR) has been less
defined yet and was addressed in the present study. We performed expression
and functional analysis of mouse tracheal epithelium using transgenic animals
and nAChR-subunit specific antagonists. Nicotine increased particle transport
speed (PTS) 2.5-fold, and this was sensitive to the general nAChR antagonist
mecamylamine. Strychnine, an antagonist at glycine receptors possesses a high
affinity to nAChR alpha10- and alpha9-subunits, dose-dependently inhibited the
nicotine-induced PTS increase whereas pretreatment with alpha-bungarotoxin
(inhibitor of alpha7, alpha9/alpha10) or dihydro-beta-erythoidine (inhibitor of
alpha3, alpha4, beta2, beta4) had no effect. Supportively, genetic deletion of
alpha7-subunit or overexpression of beta4-subunit in alpha3beta4alpha5-nAChReGFP mice had no impact on ciliary function. In all epithelial samples from
tracheas responding to nicotine, mRNA for nAChR alpha10- and beta2-subunits
was detected whereas other subunits were expressed irregularly (alpha2-alpha5,
alpha7, alpha9, beta4) or not at all (alpha6, beta3). The presence of alpha10subunit in ciliated cells of the tracheal epithelium was also confirmed at protein
level using immunohistochemistry.
In conclusion, nicotine is a powerful stimulus of tracheal particle transport, and
this effect is likely to be conferred by nAChR alpha10-subunits in an unusual
configuration.
Kategorie: Poster
Poster 65
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: Metabolic programming of embryonic cells
Autoren: Knelangen J.(1),Navarrete Santos A.(2),Kurz R.(3),Fischer
B.(1),Navarrete Santos A.(1),
Adressen:(1)Martin-Luther-Universität Halle|Institut für Anatomie und
Zellbiologie|Halle|Deutschland; email:julia@knelangen.de; (2)Martin-LutherUniversität Halle|Universitätsklinik und Poliklinik für Herz- und
Thoraxchirurgie|Halle|Deutschland; (3)Universität Leipzig|Abteilung für
molekularbiologisch-biochemische Prozesstechnik, BiotechnologischBiomedizinisches Zentrum|Leipzig|Deutschland
Abstract:
The increasing incidence of diseases like obesity or diabetes mellitus type II is a
worldwide problem. Epidemiological studies evidence that a metabolic
programming of such diseases occurs during embryonic and fetal development
and even as early as in the first days in ontogeny during embryo preimplantation
(developmental origin of health and disease, DOHaD hypothesis).
We used the cardiac differentiation of the murine embryonic carcinoma cell line
(ECC) P19 and the adipogenic differentiation of the murine embryonic stem cell
line (ESC) CGR8 to study the influence of glucose as a potential mechanism of
metabolic programming. Stem cells were cultured in media with different glucose
concentrations during the determination period for 48 hours. In P19-ECC the
frequencies of beating cardiomyocytes, measured by a Multi-electrode assay
(MEA), were significantly higher in the high glucose group compared to low
glucose. In CGR8 cells, a short-time exposure to low glucose concentrations
decreased the amount of terminally differentiated adipogenic cells by about 43%.
To identify possible epigenetic mechanisms leading to these effects, the promoter
methylation state and the expression profiles of marker genes as well as the
methylation of the histone H3 were measured by COBRA, qRT-PCR and a
methylation specific antibody, respectively.
We show the early impact of glucose on embryonic cell determination and
discuss potential epigenetic mechanisms that lead to long lasting effects in
terminally differentiated cell function. The employed cell lines, P19 ECC and
CGR8 ESC, are suitable models for metabolic programming studies, indicating
the determination stage as the most sensitive period for programming effects.
This work is supported by the German National Academic Foundation and the
German Research Council Grant NA418/4-2.
Kategorie: Poster
Poster 66
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: Rage mediates tumor-promoting effects of s100a4 in thyroid carcinoma
Autoren: Reddy-Medapati M.(1),Dahlmann M.(2),Pathak A.(3),Nason R.(3),Stein
U.(4),Hombach-Klonisch S.(5),
Adressen:(1)UoManitoba|Human Anatomy &amp; Cell
Science|Winnipeg|Canada; (2)Max-Delbrück-Center|Molecular
Medicine|Berlin|Germany; (3)UoManitoba|Surgery|Winnipeg|Canada; (4)MaxDelbrück Center|Molecular Medicine|Winnipeg|Germany; (5)UoManitoba|Human
Anatomy and Cell Science|Winnipeg|Canada; email:hombach@cc.umanitoba.ca
Abstract:
Human thyroid cancer (TC) in the most frequently occurring cancer of endocrine
glands. Some TC’s develop an aggressive phenotype with poor prognosis due to
metastases and treatment failures. The exact molecular mechanisms for this
aggressive phenotype are not well understood. Our group has previously shown
that relaxin-like peptides increase TC cell migration and in-vivo tumor growth. The
pro-migratory effects are mediated via the calcium-binding protein S100A4. We
have identified expression of the multi-ligand receptor RAGE (receptor for
advanced glycation end products) in human TC. We hypothesize that the
interaction between S100A4 and RAGE activates downstream signaling
pathways involved in promoting tumor growth and metastasis.
RT-PCR and Western analysis were performed to detect expression of S100A4
and RAGE in human TC cell lines and patient primary thyroid cells.
Immunofluorescence and immunohistochemistry were utilized to detect RAGE
protein in TC cells and patient tissues. RAGE pull-down was performed on TC
cells. Western blot detection of phosphorylated signaling intermediates was
performed following stimulation with human recombinant S100A4.
We showed expression of S100A4 and RAGE transcripts in human TC cell lines
and TC primary cells. RAGE protein was predominantly detected in TC cells at
the invasive front of tumors. Performing in vitro pull down assays we
demonstrated S100A4 binding to RAGE in TC cells. We identified MAPK
signaling pathways to be activated in TC cell lines by extracellular S100A4.
S100A4 activates key signaling pathways involved in TC cell migration and this
action is mediated by RAGE.
Kategorie: Poster
Poster 67
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: Curcumin modulates nf- b and pi-3k/akt inflammatory signaling pathways in
human tenocytes in vitro
Autoren: Busch F.(1),Buhrmann C.(1),Mobasheri A.(2),Stahlmann R.(3),Sperling
K.(4),Shakibaei m.(4),
Adressen:(1)LMU-München|Institute for anatomy|München|D; (2)University of
Nottingham|Faculty of Medicine and Health Sciences|Nottingham|UK; (3)Charite
Universitätsmedizin Berlin|Institute of Clinical Pharmacology and
Toxicology|Berlin|D; (4)LMU-München|Institute of anatomy|München|D
Abstract:
Objective:
Pro-inflammatory cytokines such as interleukin-1betta have been identified as the
main initiators of tendinopathies, stimulating inflammation, apoptosis and
extracellular matrix degradation. Pharmacological treatments for tendinitis are
restricted to the use of non-steroidal anti-inflammatory drugs (NSAIDs). Curcumin
targets the NF-kappaB signaling pathway but its potential for the treatment of
tendinitis has not been explored.
Materials and methods:
Human tenocytes were treated with interleukin-1betta to induce inflammation,
followed by co-treatment with curcumin at concentrations of 5 to 20 µM. Cultures
were evaluated by transmission electron microscopy, western blotting,
immunohistochemistry, apoptosis assays and NF-kappaB activation assays.
Results:
Curcumin inhibited IL-1betta-induced inflammation and apoptosis in tenocytes
cultures. Curcumin down-regulated the gene products involved in matrix
degradation (matrix metalloproteinases-1, 9 and -13), prostanoid production
(cyclooxygenase-2), apoptosis (Bax and activated caspase-3) and up-regulated
cell survival (Bcl-2), all known to be regulated by NF-kappaB. Further, curcumin
inhibited IL-1betta-induced NF-kappaB activation via inhibition of phosphorylation
and degradation of inhibitor of kappaB-alpha, inhibition of inhibitor of kappaBkinase activity, and inhibition of nuclear translocation of NF-kappaB. Finally, the
effects of IL-1betta were suppressed by wortmannin, suggesting a role for
phosphatidyl-inositol 3-kinase (PI-3K) pathway in IL-1betta signaling. Curcumin
inhibited IL-1betta-induced PI-3Kp85/Akt activation and its association with IKK.
Conclusions:
Our results suggest, for the first time, a potential role for curcumin in treating
tendon inflammation through modulation of the NF-kappaB signaling which
involves PI-3K/Akt.
Kategorie: Poster
Poster 68
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: Overexpression of human HMW FGF-2 but not LMW FGF-2 rescues human
corneal endothelial cell survival after lentiviral gene transfer
Autoren: Valtink M.(1),Knels L.(1),Stanke N.(2),Engelmann K.(3),Lindemann
D.(2),Funk R.(1),
Adressen:(1)TU Dresden|Anatomie|Dresden|Germany; email:monika.valtink@tudresden.de; (2)TU Dresden|Virologie|Dresden|Germany; (3)Klinikum Chemnitz
gGmbH|Augenklinik|Chemnitz|Germany
Abstract:
Basic fibroblast growth factor (FGF-2) occurs in one secreted (low molecular
weight, LMW) and four nuclear (high molecular weight, HMW) isoforms. The
secreted LMW isoform acts as a mitogen on human corneal endothelial cells
(HCEC) in vitro and as a differentiation factor in situ when administered
exogenously. Here we investigated the effect of endogenous overexpression of
each of the five known isoforms of human FGF-2 on HCEC cell survival after
lentiviral gene transfer in four different culture media.
Human corneal endothelial cells were transduced with HIV-1 pseudotyped with a
modified foamy virus glycoprotein and encoding for either the secreted LMW or
one of the four codon-optimized HMW isoforms of human FGF-2. Expression of
individual isoforms of FGF-2 was assessed by western blotting. HCEC were
cultured and infected in four different media previously described for HCEC cell
cultivation or corneal organ cultivation. Cytotoxic effect of virus infection and a
possible rescue effect by endogenous FGF-2 overexpression were determined by
means of resazurin conversion assay .
Transduction with FGF-2 carrying lentiviral vectors resulted in overexpression of
the respective isoform in all tested cell populations. Western blotting after total
cell lysis (including nuclei and other organelles) proved nuclear localization of the
transduced HMW isoforms. Overexpression of HMW FGF-2 partially abolished
the cytotoxic effect of lentiviral infection and was found to promote cell survival
under minimal nutritional conditions, while overexpression of the LMW isoform
aggravated viral cytotoxicity.
HMW FGF-2 co-expression can minimize cytotoxic effects of lentiviral
transduction.
Kategorie: Poster
Poster 69
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: Alveolar surface area and tracheal innervation in the cancer-cachectic
mouse
Autoren: Graulich T.(1),Das S.(2),Krasteva G.(1),Wessels L.(1),Kummer
W.(1),Hoefler G.(2),Mühlfeld C.(1),
Adressen:(1)Justus Liebig Universität Gießen|Anatomie und
Zellbiologie|Gießen|Deutschland; email:tilman_graulich@t-online.de;
(2)Medizinische Universität Graz|Institut für Pathologie|Graz|Österreich
Abstract:
Cancer cachexia is a complex syndrome with a significant reduction of body
weight and several systemic symptoms including respiratory dysfunction. In
rodents, calorie restriction causes loss of alveolar surface area, the so-called
nutritional emphysema. Cancer-cachectic mice show hypoinnervation of the left
ventricular myocardium. Therefore, we hypothesized that alveolar alterations and
reduced tracheal innervation are present in the cancer-cachectic mouse.
C57Bl6 mice were randomly assigned to subcutaneous injection of Lewis lung
carcinoma cells (tumor group, TG) or PBS injection (control group, CG). Mice
were sacrificed 21 days later and lungs and trachea were processed for light and
electron microscopic design-based stereology or for quantitative RT-PCR.
The alveolar surface area was similar in both groups (n=9 each; TG: 516.8±140.9
cm²; CG: 493.6±66.1 cm²). The total volume of lamellar bodies did not differ
between the groups. Quantitative expression of surfactant proteins A, B, C and D
was not different between CG and TG as shown by quantitative RT-PCR. TG
trachea showed reduced innervation per μm trachea length (n=6 each; TG:
7.3±2.3 mm/μm; CG: 14.7±2.8 mm/μm; p=0.02).
In conclusion, in contrast to weight loss due to calorie restriction, there was no
loss of alveolar surface area in cancer-cachectic mice. However, the innervation
of the trachea was reduced to approximately 50 % of control values.
Kategorie: Poster
Poster 70
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: Distribution of connexin 43 during osteoblast and osteoclast in vitro
differentiation
Autoren: Tryankowski E.(1),Cavalcanti-Adam E.(2),Glenske K.(1),Hund
R.(1),Wenisch S.(1),
Adressen:(1)University of Giessen|Department of Veterinary Surgery, c/o Institute
of Veterinary Anatomy|Giessen|Germany; (2)University of Heidelberg|Department
of Biophysical Chemistry|Heidelberg|Germany
Abstract:
Connexin 43 (cx43) is the major component of gap junctions formed between
bone cells. Its occurrence in mature osteoblasts and osteoclasts is a major
prerequisite for physiological bone remodeling. To elucidate the spatio-temporal
pattern of cx43 during cell differentiation we investigated the localization of cx43
in human mesenchymal stem cells (hMSC) and cells of the monocytemacrophage lineage.
hMSC were isolated from human spongiosa and maintained in culture.
Osteoclast-like polykaryons were generated by culturing mononuclear precursors
for 1, 7 and 10 days in standard medium supplemented with M-CSF and RANKL.
Cx43 localization was investigated by immunofluorescence, while phalloidinfluorescein-isothiocyanate was used to visualize the actin cytoskeleton.
In hMSC cx43 was scattered throughout the cytoplasm and could also be
detected along the plasma membrane. The presence of actin stress fibers was
associated with a spindle shape of the cell body. Mononuclear cells of the
monocyte-macrophage lineage revealed perinuclear localization of cx43 during
the initial stage. After one week in culture, cells were associated in clusters in
order to fuse. When polykaryons were formed, the cells showed a podosomal
patterning of the actin cytoskeleton and marked cx43 localization along the
plasma membrane.
The data presented here indicate that cx43 is already present during early stages
of cell differentiation. The signaling in hMSC is in contrast to the distribution
pattern of cx43 observed in osteoclast precursors, suggesting that the signal
observed within the region of the Golgi apparatus might be possibly due to
upregulation of the gap junctional component at the beginning of fusion.
This work is supported by the DFG/Collaborative Research Center Transregio 79.
Kategorie: Poster
Poster 71
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: Cholinergic chemosensory cells sensing the luminal microenvironment are
present in the auditory tube
Autoren: Krasteva G.(1),Hartmann P.(1),Wessels L.(1),Papadakis T.(1),Weihe
E.(2),Schütz B.(2),Ibanez-Tallon I.(3),Kummer W.(1),
Adressen:(1)Justus-Liebig-Universität Gießen|Institut für Anatomie und
Zellbiologie|Gießen|Deutschland; email:Gabriela. Krasteva@anatomie.med.unigiessen.de; (2)Philipps-Universität Marburg|Institut für Anatomie und
Zellbiologie|Marburg|Deutschland; (3)Max-Delbrück-Centrum|Molekulare
Medizin|Berlin-Buch|Deutschland
Abstract:
Recently, we have shown that brush cells of the trachea are cholinergic and affect
respiration upon stimulation with cycloheximide, a bitter tasting substance. Here,
using two different mouse strains expressing eGFP under the control of the
promoter of choline acetyltransferase (ChAT), we show the presence of identical
solitary cholinergic villin-positive brush cells also in the mouse auditory tube. They
also express the vesicular ACh transporter and proteins of the taste transduction
pathway such as alpha-gustducin, phospholipase C beta 2 (PLCbeta2), and
transient and transient receptor potential cation channel subfamily M member 5
(TRPM5). High expression was found for TRPM5 and PLCbeta2 . In general, the
observed immunoreactivity for alpha-gustducin was weak and in ca. 10% of the
brush cells even absent. Messenger RNA for bitter taste receptor (Tas2R) 105
and Tas2R108 involved in perception of the bitter substances cycloheximide and
denatonium as well as in perception of acyl-homoserine lactones, bacterial
quorum-sensing molecules, were detected in the respiratory epithelium. Using a
transgenic mouse that expresses eGFP under the control of the promotor for the
alpha3-subunit of the nicotinic ACh receptor, we identified direct contacts
between alpha3+ neuronal fibres and brush cells in the auditory tube resembling
those observed in the trachea. A subpopulation of these fibers was CGRPimmunoreactive.
We conclude that brush cells present in the auditory tube are chemosensory
cholinergic cells. Analogous to tracheal brush cells, they are equipped with all
molecules essential for sensing the composition of the luminal microenvironment
and for communication of the changes to the CNS.
Kategorie: Poster
Poster 72
Rubrik: 4.Zellbiologie
Abstract Nr.:4
Titel: Deficiency for the mitochondrial rhomboid protease parl affects
mitochondrial energy production and causes sarcopenia in postpubertal mice
Autoren: Hartmann D.(1),Deppe C.(1),Andrei-Selmer C.(1),
Adressen:(1)Bonn|Anatomie|Bonn|NRW; email:dhartman@uni-bonn.de
Abstract:
Sarcopenia, the aging – related loss of muscle mass, is a major problem of
developed countries. Morphologically, it is unrelated to neurogenic atrophy or
muscular dystrophy, featuring fiber diameter reduction as a major hallmark. The
underlying molecular pathogenesis is still debated.
Here we demonstrate that inactivation of the rhomboid protease PARL, an
apoptosis regulator situated at the mitochondrial crista neck region, causes a
similar phenotype in mice.
In knockout mice, muscle fibers terminate growth around P30 and in some
muscles even shrink, reaching a 50% loss of caliber at the time of death around
P80. Histologically, fibers appear surprisingly normal, without signs of dystrophic
fiber destruction or neurogenic atrophy. Megamitochondria are occasionally seen
in individual fibers. Unexpectedly, we did not observe any evidence for apoptosis.
Biochemically, PARL - deficient muscle fibers are characterized by a severe early
reduction of respiratory chain complex proteins SDHA / complex II and ATP
synthase B / complex V. Surprisingly, activation of the energy sensor protein
AMPK is reduced in parallel, while we did not observe a loss of mitochondrial
DNA.
Taken together, PARL defciency affects muscle fiber integrity independent of its
role in apoptosis control. Rather, its absence affects at least part of the
respiratory chain and surprisingly also intracellular energy monitoring, whereby according to fiber morphology - the changes remain below the level of severity of
typical OXPHOS mutants. This moderate impairment of mitochondrial energy
production may explain differences to OXPHOS mutants causing muscular
dystrophy and provides a novel mechanistic explanation for sarcopenia.
Kategorie: Poster
Poster 73
Rubrik: 5.Entwicklungsbiologie
Abstract Nr.:5
Titel: How urogenital sinus and wolffian ducts contribute to early human vaginal
development.
Autoren: Fritsch H.(1),Richter E.(1),Adam N.(1),
Adressen:(1)Medizinische Universität Innsbruck|Sektion Klinisch Funktionelle
Anatomie|Innsbruck|Österreich; email:helga.fritsch@i-med.ac.at
Abstract:
Introduction: Up to now the derivation of the vagina from the Muellerian ducts and
the contribution of the urogenital sinus and the Wolffian ducts still raise questions.
Cellular and molecular mechanisms involved in rodent vaginal development have
led to new ideas concerning vaginal development and have opened a large gap
between the knowledge in mice and the concepts in humans.
Materials and methods: To compare the findings in mice with human vaginal
development and to fill part of this gap we analysed the interaction of the
urogenital sinus, Wolffian ducts and Muellerian ducts in 8-14 week-old human
specimens using immunohistochemical methods. Monoclonal antibodies were
directed against cytokerartin (CK) 14, 19, Vimentin, Laminin, p63, E-Cadherin,
Caspase-3, Ki67, HOX A13 and BMP-4.
Results: Our immunohistochemical analysis reveals that in week 8-9 Muellerian
duct epithelium becomes p63 positive when p63-positive cells originating from the
sinus epithelium reach the caudal tip of the fused Muellerian ducts via Wollfian
ducts. In succession the lumen of the fused Muellerian ducts is closed by an
epithelial plug of pale and Vimentin-positive cells. Subsequently the resulting
epithelial tube enlarges by proliferation of the basal p63-positive cells. First signs
of squamous differentiation are detected in week 14 by the occurrence of CK 14positive cells.
Conclusions: According to our results all three components: urogenital sinus,
Wolffian ducts and Muellerian ducts, interact in human vaginal formation. The
sinus epithelium provides p63-positive cells, the Wollfian ducts act as
“transporter” and the Muellerian ducts build up the guiding structure for the
vaginal anlagen. Epithelial differentiation starts at the end of the period we
studied and runs in caudo-cranial direction.
Kategorie: Poster
Poster 74
Rubrik: 5.Entwicklungsbiologie
Abstract Nr.:5
Titel: Development of the lymphatic vessels in the human anorectal region
Autoren: Alessandrini P.(1),Fritsch H.(1),
Adressen:(1)Medizinische Universität Innsbruck|Anatomie Sektion für Klinisch
funktionelle Anatomie|Innsbruck|Österreich; email:petra.alessandrini@i-med.ac.at
Abstract:
Purpose: In the course of the last years different theories about the development
of the lymphatic vessels have been enunciated. Until today, however, the
anorectal region was never examined accordingly. This study aims to determine
how the development of the lymphatic vessels in this region proceeds, and,
especially, how they are distributed there. Eventually, conclusions as to the
drainage of the lymph in the anorectal region and the anal canal respectively may
be achieved.
Methods: A total of 37 fetuses between 10th week and newborn were
investigated by histological and immunohistochemical staining. The expression of
D2-40 and CD-31 markers was investigated and histologically evaluated.
Results: The first lymphatic vessels could be detected in the rectal ampulla at the
age of the 10th week, at the same time lymphatic vessels appear in the perianal
skin whereas the positive detection of lymphatic vessels with contact to the anal
canal could be demonstrated only at the beginning of the 16th week. Cells of the
anal sinus appear simultaneously with lymphatic vessels in the rectal ampulla and
also exhibit an expression of D2-40.
Conclusion: The fact that the lymphatic vessels appear simultaneous in the rectal
ampulla and the perianal skin shows that they converge upon each other.
We suppose that the epithelia of anal sinuses attract the lymphatic vessels.
Kategorie: Poster
Poster 75
Rubrik: 5.Entwicklungsbiologie
Abstract Nr.:5
Titel: Expression of c-myc during migration of myogenic precursor cells into the
limb bud of chicken embryos and validation of shrna constructs for functional
analysis
Autoren: Hellwig I.(1),Brand-Saberi B.(2),Bergmann M.(3),
Adressen:(1)Christian-Albrechts-Universität zu Kiel|Anatomisches
Institut|Kiel|Deutschland; email:i.hellwig@anat.uni-kiel.de; (2)Ruhr-Universität
Bochumg|Institut für Anatomie / Abteilung für Anatomie und Molekulare
Embryologie|Bochum|Deutschland; (3)Justus-Liebig-Universtiät Gießen|Institut
für Veterinär-Anatomie, -Histologie und -Embryologie|Gießen|Deutschland
Abstract:
Background & aims:
The proto-oncogene c-Myc promotes the progression through the cell cycle
(Amati 2001).
To address the question whether distal migration of myogenic precursor cells
within the limb bud – after delamination of cells due to de-epithelialization - is a
consequence of proliferation, signal-mediated directed migration or a mixture of
both, we analyzed the expression pattern of c-myc and designed shRNA
constructs for downregulation of c-myc.
Material and methods:
Using in situ hybridization we analyzed the expression patterns of c-myc in
chicken embryos of stages HH 17 to HH 30. To introduce the designed shRNA
constructs into the chicken embryos we applied in ovo electroporation followed by
re-incubation of 24-36h. Afterwards, we monitored c-myc mRNA expression in the
limb bud. Furthermore, we investigated Pax3 expression following c-Myc knockdown by in situ hybridization.
Results:
We could demonstrate c-myc mRNA expression in the sklerotome, the medial
and lateral lips of the dermomyotome, as well as in migrating myogenic precursor
cells and in the premuscle masses. A combination of two designed shRNA
constructs caused a silencing effect on c-myc and also a down-regulation of Pax3
mRNA expression.
Conclusions:
We observed strong expression of c-myc in myogenic precursor cells and in the
premuscle masses and we were able to silence c-myc mRNA expression by
knock-down experiments. Consequently this factor can be used in further
research studies do characterize the mechanism of propagation of myogenic
precursor cells within the limb bud mesenchyme.
Kategorie: Poster
Poster 76
Rubrik: 5.Entwicklungsbiologie
Abstract Nr.:5
Titel: Atoh8, a novel regulator of skeletal myogenesis and muscle regeneration
Autoren: Balakrishnan-Renuka A.(1),Yusuf F.(1),Patel K.(2),Otto A.(2),MorosanPuopolo G.(1),Chen J.(1),Theiss C.(1),Chankiewitz V.(1),Zoidl G.(1),Philippi
S.(3),Dai F.(3),Brand-Saberi B.(1),
Adressen:(1)Ruhr University Bochum|Institute of Anatomy, Department of
Anatomy and Molecular Embryology|Bochum|Germany; (2)University of
Reading|School of Biological Sciences|Reading|Uk; (3)University of
Freiburg|Institute of Anatomy and Cell Biology|Freiburg|Germany;
email:beate.brand-saberi@rub.de
Abstract:
Skeletal myogenesis and myogenic regeneration are essentially very similar
processes that ensure that proper functional muscle tissue is formed during
development and maintained in the course of postnatal life. Satellite cells being
the main source of resident muscle stem cells are mainly responsible for the
extensive muscle growth during late embryonic development and also for muscle
regeneration in adult life. We show that ATOH8 is expressed in the somite of
chicken embryos and silencing of ATOH8 in chicken somites perturbs skeletal
myogenesis. Furthermore, we show here for the first time that ATOH8, a bHLH
transcription factor is expressed along with Pax7 in satellite cell as well as in
skeletal myoblasts. Our results show that ATOH8 is expressed in activated
satellite cells and is downregulated as cells enter terminal differentiation.
Regenerating muscle shows an upregulated ATOH8 expression at site of injury.
Preliminary data from our protein studies show, for the first time, that cytoplasmic
ATOH8 interacts with the catalytic subunit of Calcineurin (CnA beta). Our results
are the earliest report showing the involvement of ATOH8 in embryonic
myogenesis and satellite cell differentiation. We conclude that ATOH8 is essential
for the fine regulation of the essential balance between skeletal myogenesis and
self renewal of satellite cell.
Kategorie: Poster
Poster 77
Rubrik: 5.Entwicklungsbiologie
Abstract Nr.:5
Titel: Premature formation of extraembryonic endoderm in the rabbit blastocyst
Autoren: Schwartz P.(1),Halacheva V.(1),Viebahn C.(1),
Adressen:(1)Georg-August-Universität|Zentrum Anatomie, Abteilung Anatomie
und Embryologie|Göttingen|Germany; email:cviebah@gwdg.de
Abstract:
Differentiation of the embryoblast (inner cell mass) is considered to start with the
separation into epiblast (primitive ectoderm) and hypoblast (primitive endoderm),
the latter of which is now known to be extraembryonic tissue responsible for
establishing body plan coordinates. A number of molecular markers and signals
have been described recently and were allocated to the few tissues known in
mouse embryos of that stage. In view of additional early extraembryonic tissue
types and the variety of morula and blastocyst forms amongst mammals, the
question arises which extraembryonic tissue is formed first from embryoblast in
other species than the mouse. The present study uses serial sections for highresolution light and electron microscopy on early rabbit blastocysts. After little
morphological differences exist between inner and outer blastomeres at the time
of initial blastocyst formation (4.0 days post conception, dpc), dark and slender
trophoblast cells in the enlarged blastocyst (4.5 dpc) can be distinguished from
light and round embryoblast cells, which may lie in several layers but do not show
any overt morphological differentiation. The embryoblast remains similar in
structure up to 4.75 dpc, by which time isolated or small groups of flat cells have
formed a discontinuous second cell layer on the inside of the mural trophoblast.
This appearance of extraembryonic endoderm (presumably yolk sac precursor
cells) in the absence of hypoblast differentiation is unexpected but reminiscent of
extraembryonic mesoderm appearing prior to intraembryonic mesoderm in higher
primates; correlation with expression of endoderm markers such as GATA4/6
may reveal its functional significance.
Kategorie: Poster
Poster 78
Rubrik: 5.Entwicklungsbiologie
Abstract Nr.:5
Titel: Cxcr4 mutants exhibit malformations of the perineal muscle and the genital
tubercle
Autoren: Eulitz M.(1),Rehimi R.(1),Brand-Saberi B.(1),
Adressen:(1)Ruhr-Universität Bochum|Anatomie und Molekulare
Embryologie|Bochum|Deutschland; email:mona.eulitz@rub.de
Abstract:
In many invertebrates and vertebrates, the cloaca serves as a common chamber
into which gastrointestinal and urogenital tracts converge. While birds maintain a
single cloacal opening throughout their lives, this situation only exists during
embryonic development in most mammals. In marsupials and placentals, the
common function of the cloaca is lost and two to three openings develop: anal,
urethral and vaginal in females. The distal end of the cloaca is guarded by a ring
of cloacal muscles or sphincters, the equivalent of perineal muscles in mammals.
Information about the mechanism and signaling pathways active during cloacal
muscle formation has only surfaced recently. It has been shown that
CXCR4/SDF-1 axis play an important role during cloacal muscle precursor
migration and formation.
In our study, we analyze the morphology and development of the murine genital
tubercle utilizing scanning electron microscopy (SEM). Our SEM analysis
revealed morphological malformation in the development of the cloacal/genital
tubercle in CXCR4 mutant mice compared to wild type embryos. In order to
examine whether these morphological changes are due to differences in the
perineal muscle development, we hybridized wild-type (CXCR4+/+),
heterozygous (CXCR4+/-), and homozygous (CXCR4-/-) mice embryos with a
murine probe for MyoD. We found a considerable decrease of MyoD signal in the
perineal region associated with malformation of the perineal muscles in
CXCR4+/- and CXCR4-/- mice as compared to wild type embryos. These results
show that CXCR4 signaling is critical during the development of perineal muscles
in murine embryos.
Kategorie: Poster
Poster 79
Rubrik: 6.Reproduktionsbiologie
Abstract Nr.:6
Titel: Expression of cytoskeletal proteins during folliculogenesis in the ovary of
Japanese quail (coturnix japonica)
Autoren: Rodler D.(1),Sinowatz F.(1),
Adressen:(1)Universitaet Muenchen|Department of Veterinary
Sciences|Muenchen|Deutschland; email:d.rodler@anat.vetmed.uni-muenchen.de
Abstract:
The cytoskeletal properties of the cells of the differentiating follicle wall (granulosa
cells, theca cell layers) are not known in any detail in birds. In mammalian follicles
a network of specialized cytoskeletal components has been described, which
comprises microtubuli, intermediate filaments and actin microfilaments, but in
birds the cytoskeletal machineries generating this structural organization
supporting the giant polylecithal avian oocytes have not been extensively
investigated. In the present communication we present ultrastructural and
immunohistochemical data (immunohistochemical localisation of alpha-actin,
smooth muscle actin, non-muscular myosin, tubulin and several classes of
intermediate filaments like cytokeratins, vimentin, desmin, lamin A and lamin B1)
in the follicular wall (granulosa cells and theca cells) and in the growing oocyte
during folliculogenesis in the Japanese quail (Coturnix japonica). We discuss their
putative role in follicle stability, transport of yolk components into the oocyte and
in ovulation.
Kategorie: Poster
Poster 80
Rubrik: 6.Reproduktionsbiologie
Abstract Nr.:6
Titel: Effect of ph and temperature on murine sperm motility
Autoren: Wandernoth P.(1),Mannowetz N.(2),Ruffing U.(3),Wolf A.(2),Wennemuth
G.(2),
Adressen:(1)Universität des Saarlandes|Institut für Anatomie und
Zellbiolgie|Homburg|Deutschland; email:petrawandernoth@gmx.de;
(2)Universität des Saarlandes|Institut für Anatomie und
Zellbiologie|Homburg|Deutschland; (3)Universtät des Saarlandes|Institut für
Anatomie und Zellbiologie|Homburg|Deutschland
Abstract:
The extracellular as well as the intracellular pH of sperm is important for
maturation, motility and capacitation. An acidic pH in the epididymal fluid induces
a quiescent state in sperm until ejaculation. When entering the uterus, sperm
encounter an environment with increased temperature and pH, which affects
sperm maturation. The initiation of motility and the hyperactivation phase of
capacitation depend on an elevation of internal pH.
In this project we focused on sperm motility and analyzed beat frequency under a
variety of conditions, in order to identify the direct correlation between
temperature and extracellular pH. An increase in temperature alone leads to an
increase in beat frequency from 3 Hz to 7 Hz. To characterize the influence of
extracellular pH, we incubated sperm in buffers with basic (7.8; 8.2) or acidic (7.0;
6.6) pH over a period of 3 hours. Neither beat frequency nor the intracellular pH,
determined by BCECF measurements, were affected. In a second step, in which
we manipulated the intracellular pH by means of the K+ ionophor nigericin, we
discovered that the combination of a high temperature with an elevated
intracellular pH triggered the temperature effect significantly. Under these
conditions sperm respond with an additional acceleration of beat frequency from
3 Hz to 13 Hz.
In summary, we showed that intracellular pH is independently regulated and not
linked with the extracellular pH. Furthermore, we were able to demonstrate a
direct interaction between intracellular pH and beat frequency, which establishes
the importance of pH for murine motility.
Kategorie: Poster
Poster 81
Rubrik: 6.Reproduktionsbiologie
Abstract Nr.:6
Titel: Structural modifications of the feto-placental interface in pregnancy induced
hypertension
Autoren: FRANDES C.(1),RADU A.(1),SFERDIAN M.(1),STRECU L.(1),
Adressen:(1)"VASILE GOLDIS" WESTERN UNIVERSITY OF ARAD|FACULTY
OF MEDICINE,PHARMACY AND DENTAL MEDICINE|ARAD|ROMANIA;
email:frandes.corina@gmail.com
Abstract:
INTRODUCTION Preeclampsia is a major problem of modern obstetrics and
various studies are mentioning PIH as a severe complication, one of the largest
causes of maternal and perinatal morbidity and mortality of about 5 – 7%
pregnancies throughout the world. Presenting the frequent structural
modifications of the feto-placental interface, in pregnancy induced hypertension
(PIH) .
MATERIAL AND METHOD. We have studied the main microscopical
modifications of 87 placentas obtained after delivery for two groups: one group
(Ls=57), representing parturients with PIH and another group (Lc = 30)
normotensive parturients. The samples obtained by sections were specificaly
prepared for the study of 3 types of histological stains and 4 types of
immunohistochemical stains(anti-VIM,anti-ACTIN,anti-CD34,anti-KI67). For the
histological examination we used optical microscopy for observing mainly the
lumen of spiral arteriole and changes in its tunica intima and media.
RESULTS. We registered the following structural modifications in the pregnancies
with PIH versus normal ones: changes in endothelium , fibrinoid necrosis , the
hypertrophy of tunica media , bridging syncytial knots, avascular small terminal
villi with hyaline fibrosis of the stroma, and the thrombosis of the spiral arterioles
,placental infarction and thrombosis
CONCLUSIONS. A better understanding of the immunohistological damages
demonstrated through our study, concerning the preeclamtic feto-maternal
interface, will change in the future, our understanding about the role of this
placental unit in PIH.
KEY WORDS: Pregnancy Induced Hypertension (PIH), feto-maternal interface,
placental structural modifications
Kategorie: Poster
Poster 82
Rubrik: 6.Reproduktionsbiologie
Abstract Nr.:6
Titel: Cytoarchitectonical alterations evidentiated by morphological and
histochemical methods in common placentary lesions
Autoren: FRANDES C.(1),RADU A.(1),
Adressen:(1)"VASILE GOLDIS" WESTERN UNIVERSITY OF ARAD|FACULTY
OF MEDICINE,PHARMACY AND DENTAL MEDICINE|ARAD|ROMANIA;
email:frandes.corina@gmail.com
Abstract:
Introduction: Studies regarding architectural morphological changes in placentas
in the last decade demonstrates that the placenta is not simply a transient
embryonic annex but an organ that contains a series of obscure information,
responsible for malformative pathology or prematurity. Investigation of this organ
is required at both macroscopic and especially microscopic levels in order to
differentiate the usual from the extreme cytoarchitectonical changes.
Matherial and method: Placental morphology is studied by hematoxylin-eosin
stain. The study group included 87 placentas from vaginal births with immature
fetuses showing various developmental disorders. The study material was
evaluated macroscopically and the microscopic study was performed on HE
stained histological sections and sections with PAS histochemical staining,
elective method for highlighting the fibrinoid.
Results and disscutions: The study of the selected cases in the hereby lot proves
the existence of placental lesions, which could thus provide a framework of
staging the degree of placental insufficiency according to the number of injuries
that were grouped into three degrees of severity (mild, medium and high). The
histochemical stain study shows the need for a thoroughgoing study of
microscopic methods and contemplates the using of other of maximum
accuracy(IHC).
Conclusions: The presence of placental lesions and their observation by methods
used in the hereby study suggests the intervention of the placental factor in the
pathogenetic chain of prematurity and fetal growth disorders, which requires
careful study of this transitional organ through
macro and microscopic
confounding methods.
Keywords: architectural modifications on placentas level, developmental
disorders, histochemical staining
Kategorie: Poster
Poster 83
Rubrik: 6.Reproduktionsbiologie
Abstract Nr.:6
Titel: Isolation, cultivation and identification of luteal cells in goat ovary
Autoren: Chen S.(1),Lu X.(1),Zhao S.(2),
Adressen:(1)Northwest Agriculture and Forest University|College of Veterinary
Medicine|Yangling|China; email:csl_1359@163.com; (2)University of
Freiburg|Institute of Anatomy and Cell Biology|Freiburg|Germany
Abstract:
To characterize luteal cells and differentiate them from granulose cells and
interstitial cells, Corpus luteum, follicles and connective tissue were isolated from
goat ovary and dissociated cultures were prepared respectively. Goat luteal cells
were dissociated better by using Collagenase Ⅳ than other enzymes. The
proliferation rate of granulose cells and interstitial cells was much faster than that
of luteal cells. To find out the biomarkers of luteal cells, RT-PCR for 3beta-HSD
and 17beta-HSD7 and immunocytochemistry against different markers of DNES
(diffuse Neuro-Endocrine System) were performed in above cultured different cell
types. Our results showed that mRNA of 3beta-HSD was detected in all three
kinds of cells. However, high level of mRNA for 17beta-HSD7 was found in luteal
cells, less in granulose cells, but not in interstitial cells. 3beta-HSD, P450scc, 5HT and NSE were positive in all three cell types. 17beta-HSD7 and S100 were
positive in luteal cells and granulose cells, but not in interstitial cells. OT and SYP
were positive only in luteal cells. So OT and SYP can be used as specific markers
for luteal cells. Finally, we examined telomerase activity in all cell types.
Telomerase activity was detected in granulose cells and interstitial cells, but not in
luteal cells. Conclusion: SYP and OT by IHC can be used as biomarkers for
identification of luteal cells in goat ovary. Telomerase activity is not able to be
detected in luteal cells, indicating that these cells lost mitosis activity.
Kategorie: Poster
Poster 84
Rubrik: 6.Reproduktionsbiologie
Abstract Nr.:6
Titel: Localization of ck2 subunits and its possible binding partners tnp1 and kif5c
in the murine testis
Autoren: Mannowetz N.(1),Wennemuth G.(1),Kartarius S.(2),Montenarh M.(2),
Adressen:(1)Universität des Saarlandes|Institut für Anatomie und
Zellbiologie|Homburg|Deutschland; email:nadja.mannowetz@uniklinikumsaarland.de; (2)Universität des Saarlandes|Institut für medizinische Biochemie
und Molekularbiologie|Homburg|Deutschland
Abstract:
Protein kinase CK2 regulates a variety of cellular processes, such as
gametogenesis. The holoenzyme is a heterotetramer consisting of two regulatory
beta-subunits and the two catalytic subunits alpha and alpha'. The individual
subunits have been shown to bind to an abundance of cellular proteins in various
cell types, i. e. the transition nuclear protein 1 (TNP1). It has been demonstrated
that expression of both TNP1 and CK2 peaks in pachytene spermatocytes and
round spermatids. In neurons, CK2alpha' interacts with the neuronal motor
protein KIF5C, which, in turn, has been shown to be expressed also in
spermatids. With regard to a possible interaction between CK2 and TNP1 and
KIF5C respectively, we asked here for the localization of CK2 subunits, TNP1 and
KIF5C
in
spermatogenic
cells
and
mature
spermatozoa.
With
immunohistochemistry we found CK2beta and TNP1 to be present in cells of
early spermatogenesis, whereas CK2alpha, CK2alpha' and KIF5C were localized
in late spermatogenic cells. Immunofluorescence with epididymal sperm showed
CK2alpha-, TNP1- and KIF5C-specific signals in the acrosome, whereas CK2beta
and CK2alpha' were additionally present in the midpiece. Immunogold labeling
revealed a subcellular localization of CK2alpha and KIF5C in the developing
acrosome. However, for CK2beta, CK2alpha' and TNP1 no or only faint signals
restricted to the midpiece were visible. Although we so far did not directly
demonstrate interactions between CK2 proteins and TNP1 and KIF5C
respectively, we conclude that TNP1 and KIF5C are possible binding partners for
CK2 thereby regulating both spermatogenesis and sperm functions.
Kategorie: Poster
Poster 85
Rubrik: 6.Reproduktionsbiologie
Abstract Nr.:6
Titel: Pde expression and pde-dependent contractility of seminiferous tubules of
the testis.
Autoren: Feuerstacke C.(1),Mietens(2),Tasch(2),König(3),Middendorff(2),
Adressen:(1)Justus Liebig Universität Giessen|Anatomie und
Zellbiologie|Giesse|Deutschland;
email:Caroline.Feuerstacke@anatomie.med.uni-giessen.de; (2)Justus Liebig
Universität Giessen|Anatomie und Zellbiologie|Giessen|Deutschland;
email:Andrea.Mietens@anatomie.med.uni-giessen.de; (3)Universität
Lübeck|Institut für Anatomie|Lübeck|Deutschland;
email:Ralf.Middendorff@anatomie.med.uni-giessen.de
Abstract:
In the human testis, myofibroblasts are the main cellular components of the
lamina propria (LP) of seminiferous tubules. These cells are crucial for the
transport of immature sperm towards the epididymis. The second messenger
cGMP contributes to the regulation of contractile cell function. Duration of cGMP
action depends on different phosphodiesterases (PDEs) hydrolyzing cGMP. The
PDE5A inhibitor sildenafil is used in an increasing number of young patients to
treat pulmonary hypertension.
Laser capture microdissection combined with RT-PCR was used to characterize
PDE expression. Functional aspects were investigated by newly developed
collagen gel-based assays in combination with video microscopy. All 22 PDE
isoforms were detected in the human testis. By use of additional cell type-specific
primer pairs and comparison to Sertoli cell only syndrome. PDE1A, PDE1B,
PDE1C, PDE5A, PDE3B, PDE9A and PDE10A could be localized to the LP,
whereas PDE2A, PDE3A and PDE11A were absent. Different to the PDEs
mentioned before PDE1C was lacking in thickened LP nearly regularly found in
disturbed spermatogenesis. Involvement of PDE5A in contractility of isolated
seminiferous tubules was revealed by our new functional assays.
The specific expression profile of PDEs in the LP of the human testis and effects
of PDE5 inhibition on contractility in seminiferous tubules point to a well
orchestrated functional interplay of PDEs for testicular sperm transport. Absence
of PDE1C in thickened LP might hint on a proliferation deficit of contractile cells in
male infertility suggesting a new therapeutical target.
Kategorie: Poster
Poster 86
Rubrik: 7.Immunbiologie
Abstract Nr.:7
Titel: The immunopathology of MP4-induced experimental autoimmune
encephalomyelitis is complement-dependent
Autoren: Hundgeburth L.(1),Pauly R.(1),Rottlaender A.(1),Rodi M.(1),Addicks
K.(1),Kuerten S.(1),
Adressen:(1)Köln|Institut I für Anatomie|Köln|Deutschland;
email:stefanie.kuerten@uk-koeln.de
Abstract:
The contribution of the complement system to the pathogenesis of multiple
sclerosis (MS) has remained unclear. So far, studies of experimental autoimmune
encephalomyelitis (EAE) – the animal model for MS – were largely hampered by
the absence of a pathogenic B cell/antibody response in most models. We have
previously demonstrated that the myelin basic protein (MBP)-proteolipid protein
(PLP) fusion protein MP4-induced EAE is B cell- and antibody-dependent. Here
we show that complement is needed for MP4-specific antibodies to trigger CNS
pathology. Demyelinated lesions in the CNS were colocalized with complement
and antibody depositions. In addition, B cell deficient JHT mice reconstituted with
MP4-reactive serum showed significantly attenuated clinical and histological EAE
after depletion of complement by injection of cobra venom factor (CVF). Our data
also suggest that the complement system is critically involved in the generation of
the MP4-specific B cell response, while the T cell response was unaffected in
complement-depleted mice. In conclusion, we propose that the MP4 model may
serve as a valuable tool for differential downstream mechanistic studies of
complement-mediated mechanisms in the pathogenesis of MS.
Kategorie: Poster
Poster 87
Rubrik: 7.Immunbiologie
Abstract Nr.:7
Titel: Histopathological and immunohistochemical study of two pilomatrixoma
cases
Autoren: Anca I.(1),Cianga C.(2),Lucian Laurentiu I.(1),Gabriela Florenta
D.(3),Petru C.(2),
Adressen:(1)"Gr.T. Popa" University of Medicine and Pharmacy|Anatomy
Institute|Iasi|Romania; email:anca_indrei@yahoo.com; (2)"Gr.T. Popa" University
of Medicine and Pharmacy|Immunology|Iasi|Romania; (3)"Gr.T. Popa" University
of Medicine and Pharmacy|Pathology|Iasi|Romania
Abstract:
Pilomatrixoma, a rare hair follicle tumor, originates and develops from the
follicular matrix cells. We present below two pilomatrixoma cases in two patients
aged 3 and 9 respectively. The purpose of this study consisted in the
identification of the tumor cells’ nature and the assessment of their growth rate,
as well as the determination of its associated inflammatory reaction. Materials
and methods: In both situations, the clinical examination revealed non-ulcerated,
clearly shaped and painful tumors on the upper eyelid that led to the diagnosis of
dermoid cyst. After the surgical removal of tumors, the tissues were fixed and
paraffin embedded. Some of the tissue sections were stained with hematoxylin,
eosin and PAS, and others were subjected to the immunohistochemical
examination for the purpose of determining the expression of certain molecules of
interest: cytokeratins, CD3, CD20, CD68, PCNA. Results: the research
highlighted that the tumors, well delimitated by conjunctive capsules, are formed
by irregular islands of epithelial cells specific to pilomatrixoma: basaloid,
transitional and ghost cells, differently represented in the two cases. The
transitional and basaloid tumor cells of the 3 years old patient were well
represented, 15% of them being PCNA+. In the patient aged 9, the basaloid and
transitional cells are almost absent, the entire tumor architecture being dominated
by ghost and giant cells, CD68+. Conclusions: the histopathological analysis is
essential
and
sufficient
for
diagnosing
the
pilomatrixoma.
The
immunohistochemistry provides additional information that is extremely important
for interpreting the lesion stage and for formulating the prognostic.
Kategorie: Poster
Poster 88
Rubrik: 7.Immunbiologie
Abstract Nr.:7
Titel: Transport of bacteria and cell-type specific induction of signaling in the
tracheal airway epithelium of mice
Autoren: Bermbach S.(1),Rupp J.(2),König P.(1),
Adressen:(1)Universität zu Lübeck|Institut für Anatomie|Lübeck|Deutschland;
(2)Universität zu Lübeck/Universitätsklinikum Schleswig-Holstein|Institut für
Medizinische Mikrobiologie und Hygiene|Lübeck|Deutschland;
email:koenig@anat.uni-luebeck.de
Abstract:
The airway epithelium is constantly in contact with inhaled bacteria without
inducing inflammation. However, sometimes this barrier is broken and infection
occurs. To better understand the cellular and molecular interaction of bacteria
and airway epithelium we used a short term culture model of the murine trachea.
Direct interactions between bacteria and the airway epithelium were analyzed
using high speed video microscopy. Downstream signaling events in the
interaction of bacteria with the epithelium were determined on a molecular level
by real time RT-PCR of proinflammatory cytokine mRNA. To characterize celltype specificity of the bacteria-induced immune response, nuclear NFkappaB
translocation was assessed using immunohistochemistry and confocal
microscopy. Despite the lack of a continuous mucus layer, bacteria were
effectively transported by direct interaction with cilia and cilia-driven fluid flow,
preventing prolonged interaction of airway epithelial cells with bacteria. In line
with this observation, incubation with S. aureus failed to induce an increase of
mRNA levels for proinflammatory cytokines in the epithelium. However,
incubation of the trachea with H. influenzae increased respective mRNA levels
but NFkappaB translocation was only detected in non-ciliated cells. The selective
translocation of NFkappaB in non-ciliated cells was also observed after incubation
of the trachea with bacterial LPS. These observations indicate that ciliated cells
effectively transport bacteria in the absence of mucus and this interaction does
not result in NFkappaB-dependent epithelial signaling. This pathway is initiated
only in non-ciliated cells, presumably via bacterial cell wall components.
Kategorie: Poster
Poster 89
Rubrik: 7.Immunbiologie
Abstract Nr.:7
Titel: Targeting of gliadin peptide á31-49 to late endosomes of enterocytes of
infantile mice by conjugation to cholera toxin b subunit (ctb)
Autoren: Schmitz M.(1),Melcher C.(1),Sauerland C.(2),Hildebrand R.(1),Zimmer
K.(3),
Adressen:(1)Westfälische Wilhelmsuniversität Münster|Institut für
Anatomie|Münster|Deutschland; email:martina.schmitz@ukmuenster.de;
(2)Westfälische Wilhelmsuniversität Münster|Institut für Biometrie und klinische
Forschung|Münster|Deutschland; (3)Universitätsklinikum Gießen und
Marburg|Allgemeine Pädiatrie und Neonatologie|Gießen|Deutschland
Abstract:
Background: Coeliac disease (CD) - a multisystemic autoimmune inflammation of
the small intestine - is a result of a breakdown in oral tolerance to wheat gluten
and related cereals in HLA-DQ2/8 positive individuals. The translocation of
luminal antigen to LAMP- and HLA-DR-positive late endosomes of small intestinal
enterocytes is thought to be a decisive mechanism for the induction of oral
tolerance.
Methods: After intraoperative application in defined segments of the jejunum and
ileum of gliadin-naive 10-day-old mice we quantified morphometrically the
subcellular localization of both toxic and non-toxic gliadin peptides in enterocytes
by using an immunogold technique on thin frozen sections. In addition we tried to
modulate the sorting of the toxic peptide M1 (alpha31-49) towards late
endosomes based on the model of the non-toxic peptide M3 (alpha229-246) by
conjugation to cholera toxin B subunit (CTB).
Results: Unlike the control peptide M3 the toxic gliadin peptide M1 (alpha31-49)
fails to reach late endosomes of jejunal enterocytes and thereby escapes antigen
presentation by HLA-DR molecules at the basolateral membrane. Strikingly M1
being linked to cholera toxin B subunit can be rerouted to late endosomes of
enterocytes.
Conclusion: The separate pathway of gliadin peptide M1 and its absence in late
endosomes seems to be a crucial process and could explain the peptide’s
toxicity2,3 and its failure to induce T lymphocytes. Hence, the successful sorting
of M1 in late endosomes by conjugation to CTB could be a potentially promising
both preventive and therapeutic approach in the treatment of CD.
Kategorie: Poster
Poster 90
Rubrik: 7.Immunbiologie
Abstract Nr.:7
Titel: Bruton's tyrosine kinase is an important signaling molecule for toll-like
receptor-induced immunoregulatory functions in macrophages
Autoren: Vijayan V.(1),Immenschuh S.(2),Qian G.(1),Baumgart-Vogt E.(1),
Adressen:(1)Justus-Liebig University|Institute for Anatomy and Cell Biology
II|Giessen|Germany; (2)Hannover Medical School|Institute of Transfusion
Medicine|Hannover|Germany; email:Eveline.Baumgart-Vogt@anatomie.med.unigiessen.de
Abstract:
Bruton's tyrosine kinase (Btk), the gene of which is mutated in the human
immunodeficiency X-linked agammaglobulinemia, is crucial for B-cell maturation
and function. Recent evidences suggest an important role for Btk in macrophage
effector functions such as phagocytosis, cytokine production and survival . In the
current study we demonstrate that, Btk is involved in the toll-like receptor (TLR)induced gene expression of Heme oxygenase(HO)-1, which is a protein that has
been shown to have potent cytoprotective and antioxidant functions in various
inflammatory disorders. Inhibition of Btk with LFM-A13 abolished the upregulation of HO-1 by the classical TLR4 ligand LPS in realtime RT-PCR, western
blot and immunofluorescence studies with primary mouse alveolar and
RAW264.7 macrophages. Moreover, LPS-dependent induction of HO-1 gene
expression was diminished in alveolar macrophages from Btk-/- mice. This Btkdependent up-regulation of HO-1 was found to be mediated by the transcription
factor Nrf2, which is a master regulator of the antioxidant cellular defense.
Accordingly, nuclear translocation and activity of Nrf2 in LPS-stimulated
macrophages was reduced by Btk inhibition as indicated by immunofluorescence
studies and luciferase reporter gene assays. As shown by FACS analysis, the
generation of reactive oxygen species (ROS) was involved in this regulatory
pathway. In addition, TLR2, TLR7 and TLR9 ligands also induced the expression
of HO-1 in a Btk-dependent fashion suggesting Btk to have a widespread role in
TLR signaling.
Kategorie: Poster
Poster 91
Rubrik: 7. Immunbiologie
Abstract Nr.:
Titel: Tumor necrosis factor alpha (tnf-alpha) mediates cns pathology in myelin
oligodendrocyte glycoprotein-induced experimental autoimmune
encephalomyelitis
Autoren: Holland F.(1),Thomalla F.(2),Recks M.(2),Batoulis H.(2),Addicks
K.(2),Kuerten S.(2),
Adressen:(1)Köln|Institut I für Anatomie|Köln|Deutschland; (2)Köln|Institut I für
Anatomie|Köln|Deutschland; email:stefanie.kuerten@uk-koeln.de
Abstract:
Myelin-reactive CD4+ T cells of the TH1 or TH17 effector lineage are considered
to be involved in the immunopathology of experimental autoimmune
encephalomyelitis (EAE), the animal model of multiple sclerosis (MS). In myelin
oligodendrocyte glycoprotein (MOG) peptide 35-55-induced EAE of C57BL/6
mice we demonstrate that the numbers of MOG:35-55-specific TH1 and TH17
cells measured in the blood by ELISPOT before disease onset correlate with the
subsequent disease severity at the initial peak of the disease. However, once
EAE has developed, disease severity does not correlate with the antigen-specific
TH1/TH17 response, but rather with the degree of macrophage infiltration and the
amount of TNF-alpha present in the CNS. Administration of anti-TNF-alpha
antibody every other day over a period of 20 days post immunization led to
significant disease amelioration. TNF-alpha could either mediate CNS pathology
by the induction of apoptosis or via supporting inflammation, the release of
oxygen species and glutamate excitoxicity. In addition, TNF-alpha could assert
direct cytotoxic effects on oligodendrocytes and axons. Since TNF blockade using
TNFR-Fc antibody did not attenuate disease, the results indicate that a direct
inflammatory and/or cytotoxic effect of TNF-alpha seems to be involved in
mediating CNS pathology in MOG peptide-induced EAE. In conclusion, we
suggest that while CD4+ T cells are important for disease initiation, innate
immune mechanisms may be associated with disease maintenance and therefore
need to be carefully considered when evaluating therapeutic strategies and
efficacy.
Kategorie: Poster
Poster 92
Rubrik: 8.Neuroregeneration/Neurodegeneration
Abstract Nr.:8
Titel: Characterizing the role of fus and mutated fus in neurons and synaptic
contacts
Autoren: Putz S.(1),Heinrich J.(1),Schmeißer M.(1),Schmidt T.(1),Pröpper
C.(1),Ludolph A.(2),Böckers T.(1),
Adressen:(1)Universitätsklinikum Ulm|Institut für Anatomie und
Zellbiologie|Ulm|Deuschland; (2)Universitätsklinikum Ulm|Klinik für
Neurologie|Ulm|Deuschland
Abstract:
The ALS related protein FUS (Fused in Sarcoma) is a multifunctional RNA
binding protein, which is involved in multiple steps of RNA processing such as
splicing, translation and transport. Recently it has been demonstrated that FUS
mutations are causative for motor neuron degeneration in patients suffering from
amyotrophic lateral sclerosis. Beside its localization in the nucleus, FUS shows
also a somatodendritic localization in neurons and is found to translocate into
synaptic spines upon excitatory synaptic input. Based on the described putative
role of FUS in mechanisms related to plasticity and local rearrangement of
synaptic contacts we started to analyze the FUS protein with respect to its role in
dendrites and spines. Another research interest is the characterization of the
known and novel FUS mutations in cell lines and primary neuronal cultures.
Our transfection experiments with mutated FUS in cell lines and neurons revealed
a clear mislocalization of the FUS protein in the cytoplasm. Thereby the degree of
mislocalization is correlated to the aggressiveness of the disease course. In
subsequent experiments we found that endogenous FUS is present in synaptic
junctions and within the postsynaptic density. Furthermore we found a strong
association of the FUS protein with the cytoskeleton which supports the idea of
an involvement of the protein in dendritic transport processes. Taken together our
first data are supportive for the hypothesis that FUS function in neurons is
extended compared to other cell types and that the synaptic compartment is most
likely one of the sites of action
Kategorie: Poster
Poster 93
Rubrik: 8.Neuroregeneration/Neurodegeneration
Abstract Nr.:8
Titel The olfactomedin proteins pancortin and myocilin modulate apoptosis of
retinal neurons after damage
Autoren: Koch M.(1),Rosenhammer B.(1),Braunger B.(1),Paper W.(1),Tamm
E.(1),
Adressen:(1)Universität Regensburg|Lehrstuhl für Humananatomie und
Embryologie|Regensburg|Deutschland; email:Marcus.Koch@VKL.UniRegensburg.de
Abstract:
Purpose: Pancortin and Myocilin are secreted matricellular proteins that share an
olfactomedin domain. Both are modulators of non-canonical Wnt-signaling and
are found in the retina. As Wnt-signaling has been shown to protect retinal
ganglion cells (RGC) from apoptosis after damage, we investigated, if retinal
neurons of mice that are each deficient for pancortin or myocilin, respectively,
differ in their responses to injury.
Methods: Apoptosis of RGC was induced by NMDA-injection and excitotoxicity,
while apoptosis of photoreceptors was induced by light damage, a model of
retinal degeneration in rodents. Neuronal cell death was analyzed by TUNELlabeling, and quantitative morphometry, such as analyzing thickness of the outer
nuclear layer (ONL) and the number of axons in the optic nerve.
Results: RGC of pancortin mutant mice do not differ in their response to
excitotoxicity when compared to wild-type littermates. Still, in the ONL of
pancortin mutant mice, a significantly higher rate of photoreceptor loss and a
significantly higher number of TUNEL-positive cells was observed after light
damage. In contrast, photoreceptors of myocilin-deficient mice were protected
against light damage and showed less cell loss in the ONL and fewer TUNELlabeled cells. Comparable findings were observed for RGC of myocilin-deficient
mice after NMDA-induced excitotoxic damage.
Conclusions: Both pancortin and myocilin modulate apoptosis of retinal neurons
after damage, but have each different and opposite effects on the rate of RGC
and/or photoreceptor apoptosis.
Supported by DFG Forschergruppe 1075
Kategorie: Poster
Poster 94
Rubrik: 8.Neuroregeneration/Neurodegeneration
Abstract Nr.:8
Titel: Phosphorylation dependent translocation of the small heat shock proteins
hspb1/hsp25 and hspb5/ b-crystallin in neurons
Autoren: Schmidt, T.(1),Bartelt-Kirbach B.(2),Prof. Dr. Golenhofen N.(1),
Adressen:(1)Ulm|Anatomie und Zellbiologie|Ulm|Deutschland; (2)Ulm|Anatomie
und Zellbiologie|Ulm|Deuschland
Abstract:
Heat shock proteins play a major role in the development of stress tolerance.
They are upregulated upon various stresses and help the cell to survive under
pathophysiological conditions. Eleven small heat shock proteins (sHsps) are
known in mammals, some of which have neuroprotective functions or play a role
in neurodegenerative diseases. They exert the cytoprotective function via their
chaperone like activity and specific other functions such as interaction with
cytoskeletal proteins or inhibiting apoptosis. These specific functions are thought
to be regulated by phosphorylation. Since little is known about the functional role
and molecular targets of phosphorylated sHsps in neurons we studied the
subcellular distribution of two family members, i.e. HspB1 and HspB5, and their
phosphorylated forms in cultured hippocampal neurons by immunocytochemistry.
Using antibodies detecting non-phosphorylated and phosphorylated forms of
these proteins we found HspB1 and B5 mainly localized in the perikaryon and
nucleus of neurons. However, antibodies specific for the various phosphorylation
sites show additionally immunosignals in neuronal processes demonstrating
recruitment of HspB1 and B5 to neuronal processes by phosphorylation.
pHspB1(S15) and pHspB1(Ser86) localized to dendrites and especially synaptic
sites whereas pHspB5(S19) and pHspB5(S45) showed a filamentous like staining
pattern in dendrites and axons and pHspB5(S59) was found along the plasma
membrane and dendritic spines. Subcellular fractionation of rat brain lysates
confirmed localization of HspB1 and B5 in nuclear, cytosolic and synaptic
fractions. These data hint to different molecular targets of the various
phosphorylated forms of HspB1 and B5 in neurons.
Kategorie: Poster
Poster 95
Rubrik: 8. Neuroregeneration/Neurodegeneration
Abstract Nr.:
Titel: Effects of intrastriatal botulinum neurotoxin a in rats
Autoren: Wree A.(1),Holzmann C.(2),Hawltischka A.(1),Mix E.(3),Benecke
R.(3),Dräger D.(1),
Adressen:(1)Universität Rostock|Institut für Anatomie|Rostock|Deutschland;
email:andreas.wree@med.uni-rostock.de; (2)Universität Rostock|Kinder- und
Jugendklinik, Abt. Medizinische Genetik|Rostock|Deutschland; (3)Universität
Rostock|Klinik und Poliklinik für Neurologie|Rostock|Deutschland
Abstract:
Purpose: Intrastriatal cholinergic hyperactivity is responsible for motor symptoms
of Parkinson\'s disease. Studies showed that the pathological rotations inducible
by apomorphine in the Hemiparkinson animal model were inhibited after
intrastriatal injection of botulinum neurotoxin A (BoNT-A) for at least 6 months. As
a cholinergic hypoactivity is also associated with cognitive disorders the aim of
the study is to examine effects of bilateral intrastriatal BoNT-A application in rats.
Methods: 55 Rats were divided into 3 groups: animals which received bilateral
intrastriatal per 1 ng BoNT-A, which received vehicles injections, and untreated
animals. 5 weeks after BoNT- or vehicle application rats were tested by open field
(OF), elevated plus maze (EPM), accelerod (AT), water maze (WM) and radial
maze (RM) tests. Then, serial brain slices were evaluated by quantitative
histology and immunocytochemisty.
Results: BoNT-animals show significantly reduced locomotion and anxiety
behavior in OF and EPM. In addition, intrastriatal injected BoNT-A causes
impairment of balance and coordination (AT). There are no significant differences
of spatial memory (WM). Concerning long-term and working memory BoNT-and
sham-BoNT animals do not differ. The controls perform better; thus the
stereotactic operation influence animals´ behavior. Immunocytochemically there
are no significant changes in ChAT-ir neurons in the BoNT injected striata.
Conclusions: The bilateral intrastriatal application of 1ng BoNT-A affected the
balance and coordination performance. It has no significant influence on the
cognitive performance and reduces the anxiety behavior. Thus, Parkinsonism
patients with neuropsychiatric symptoms could benefit from a central application
of BoNT.
Kategorie: Poster
Poster 96
Rubrik: 8. Neuroregeneration/Neurodegeneration
Abstract Nr.:
Titel: Cognitive impairment and cerebral atrophy with IgA antibodies to NMDA
receptor
Autoren: Höltje M.(1),Maier N.(2),Gomez A.(3),Burchert R.(4),Harms L.(5),AhnertHilger G.(6),Schmitz D.(7),Wandinger K.(8),Prüss H.(5),
Adressen:(1)Charité-Universitätsmedizin Berlin|Centrum für
Anatomie|Berlin|Germany; email:markus.hoeltje@charite.de; (2)Charité University
Medicine, Berlin|Neuroscience Research Center|Berlin|Germany; (3)CharitéUniversity Medicine Berlin|Department of Psychiatry|Berlin|Germany; (4)CharitéUniversity Medicine Berlin|Department of Nuclear Medicine|Berlin|Germany;
(5)Charité-University Medicine Berlin|Department of Neurology|Berlin|Germany;
(6)Charité-University Medicine Berlin|Centrum für Anatomie|Berlin|Germany;
(7)Charité-University Medicine Berlin|Neuroscience Research
Center|Berlin|Germany; (8)Euroimmun AG|Institute for Experimental
Immunology|Lübeck|Germany
Abstract:
Dysfunction of memory, behavior, attention and planning can result from
autoimmunity to ion channels or receptors in the brain. In this study we report on
the novel association of NMDA receptor IgA (but not IgG) antibodies in patients
with progressive anterograde memory deficits resembling a primary degenerative
disorder. When applied to cultures of primary hippocampal neurons, patients’
serum but not serum from control individuals caused a strong decrease of the
levels of NMDAR and other synaptic proteins such as synapsin and vesicular
transmitter transporters. This phenomenon was accompanied by prominent
changes in NMDAR-mediated currents as detected by electrophysiology. After
removal of patient’s serum from the media these effects were reversed and
correlated with antibody titers. Immunotherapy resulted in clinical improvement
and partial regain of brain metabolism in regions of high NMDAR density.
This novel association of serum NMDAR autoantibodies of the IgA class with
neurodegeneration, and the effects of patients’ antibodies on synaptic proteins
suggest that humoral autoimmunity plays a role in the cognitive and behavioral
dysfunction of some patients. These findings provide an opportunity to treat these
patients with immunotherapy.
Kategorie: Poster
Poster 97
Rubrik: 8. Neuroregeneration/Neurodegeneration
Abstract Nr.:
Titel: Effect of the advanced glycation end products-inducer glyoxal and of
neuroprotective substances in mouse retinal explant culture
Autoren: Knels L.(1),Wurm A.(2),Valtink M.(2),Röhlecke C.(2),Ader M.(3),Funk
R.(2),
Adressen:(1)Technische Universität Dresden|Institut für
Anatomie|Dresden|Germany; email:lilla.knels@tu-dresden.de; (2)Technischen
Univerität Dresden|Institut für Anatomie|Dresden|Germany; (3)Technischen
Univerität Dresden|DFG-Center for Regenerative Therapies
Dresden|Dresden|Germany
Abstract:
AGEs can augment the risk of AMD or diabetic retinopathy. To induce AGE
formation in retinal wholemounts and to determine which retinal cell types are
predominantly affected, which pathways of cell death are involved, and if
memantine or erythropoietin can act as neuroprotectants. C57/BL6 wild type
retinal explants were placed on well inserts with photoreceptors facing the
membrane, and cultured for up to 5d with medium containing glyoxal to induce
AGE formation, and with or without memantine or erythropoietin. Control explants
were cultured without glyoxal. Samples were analysed by immunoblotting for N(epsilon)-carboxymethyllysine (CML), apoptosis inducing factor (AIF), and Bcl2associated protein X (Bax), and by RT-PCR for heme oxygenase-1 (HO-1), Bax,
AIF, and receptor for AGEs (RAGE), and immunostaining for CML and Bax.
Mouse retinal explants maintained their architecture during the cultivation period.
Treatment with glyoxal induced CML formation, increased AIF and Bax protein
levels, and up-regulated expression of AIF, Bax and HO-1. RAGE expression was
not affected. Immunostaining revealed strong CML formation mainly in the outer
nuclear layer (ONL), and Bax positivity mainly in the ONL, inner nuclear layer and
to a lesser degree in the ganglion cell layer (GCL). Memantine-treated cells had
decreased Bax protein levels, but CML amounts remained constant.
Erythropoietin showed no effect on Bax or CML production.
Glyoxal leads to formation of AGEs and in consequence to an up-regulation of
apoptotic markers in retina in vitro. While memantine may attenuate apoptotic
reactions, erythropoietin is unable to proptect the cells from eventual apoptosis.
Kategorie: Poster
Poster 98
Rubrik: 8. Neuroregeneration/Neurodegeneration
Abstract Nr.:
Titel: Differential contribution of spinal cord white and gray matter pathology to
experimental autoimmune encephalomyelitis
Autoren: Kuerten S.(1),Recks M.(1),Stormanns E.(1),Gruppe T.(1),Addicks K.(1),
Adressen:(1)Köln|Institut I für Anatomie|Köln|Deutschland;
email:stefanie.kuerten@uk-koeln.de
Abstract:
Experimental autoimmune encephalomyelitis (EAE) has been studied for decades
as an animal model for human multiple sclerosis (MS). Here we performed
ultrastructural analysis of pyramidal tract and motor neuron pathology in myelin
oligodendrocyte glycoprotein (MOG) peptide 35-55- and MP4-induced EAE of
C57BL/6 mice. Both models were clinically characterized by ascending paralysis.
Our data show that pyramidal tract and motor neuron pathology differentially
contributed to the disease. In MOG peptide EAE pyramidal fiber tract changes
prevailed, while the MP4 model additionally encompassed severe motor neuron
degeneration, which included rough endoplasmic reticulum alterations, the
presence of intracytoplasmic vacuoles and nuclear dissolution. In addition, both
models displayed distinct features of axonal damage that covered mitochondrial
swelling, decrease in nearest neighbor neurofilament distance (NNND) and an
increase of the oligodendroglial cytoplasm inner tongue formation. The extent of
pyramidal tract and motor neuron pathology was reflective of the severity of
clinical EAE in MOG peptide- and MP4-elicited EAE. Differential targeting of CNS
gray and white matter are typical features of MS pathology. The MOG:35-55 and
MP4 model may thus be valuable tools for downstream studies of the
mechanisms underlying these morphological disease correlates.
Kategorie: Poster
Poster 99
Rubrik: 8. Neuroregeneration/Neurodegeneration
Abstract Nr.:
Titel: Spinal cord histopathology of mog peptide 35–55-induced experimental
autoimmune encephalomyelitis is time- and score-dependent
Autoren: Recks M.(1),Addicks K.(1),Kuerten S.(1),
Adressen:(1)Köln|Institut I für Anatomie|Köln|Deutschland;
email:stefanie.kuerten@uk-koeln.de
Abstract:
Experimental autoimmune encephalomyelitis (EAE) is the most widely used
animal model of the human disease multiple sclerosis (MS), which is
characterized by multicentric CNS inflammation and neurodegeneration. Myelin
oligodendrocyte glycoprotein (MOG) peptide 35–55-induced EAE in C57BL/6
mice is one of the traditional EAE models. While the inflammatory component of
MOG peptide 35–55-induced (MOG:35–55) EAE has been investigated in detail,
the role of neurodegenerative processes has not yet been thoroughly
characterized. Neurodegeneration, however, is a major hallmark of MS pathology
and considered to be the morphological correlate for irreversible functional
deficits in patients. For EAE induction, mice were immunized with MOG:35–55. In
the acute, chronic and long-term chronic stage of EAE, three transverse
segments of the lumbar spinal cord were obtained, embedded in epon, stained
with methylene blue and evaluated by light microscopy. Assessment of the semithin sections showed, on the one hand, a time-dependence of spinal cord
histopathology with a transition from acute inflammation to chronic
neurodegeneration. Distinct criteria to distinguish between acute and chronic
stages of EAE were detected. On the other hand, we found a strong correlation
between the extent of white matter plaque formation and clinical disease severity.
Additionally, differences in the involvement of the anterolateral, posterior and
pyramidal tract were evident. In sum, our results delineate that MOG:35–55induced EAE is a well-suited MS model in such a way that it closely mirrors the
hallmarks of MS pathology.
Kategorie: Poster
Poster 100
Rubrik: 8. Neuroregeneration/Neurodegeneration
Abstract Nr.:
Titel: Brainstem nuclei associated with perineuronal nets are not protected
against tau pathology in progressive supranuclear palsy (psp)
Autoren: Horn A.(1),Messoudi A.(2),Roeber S.(3),Ogunlade V.(3),Kretzschmar
H.(3),
Adressen:(1)LMU Muenchen|Anatomisches Institut, Lehrstuhl I; Integriertes
Forschungs- und Behandlungszentrum für Schwindel, Gleichgewichts- und
Augenbewegungsstörungen|Muenchen|Deutschland;
email:Anja.Bochtler@med.uni-muenchen.de; (2)LMU Muenchen|Anatomisches
Institut, Lehrstuhl I|Muenchen|Deutschland; (3)LMU Muenchen|Institut für
Neuropathologie; Integriertes Forschungs- und Behandlungszentrum für
Schwindel, Gleichgewichts- und
Augenbewegungsstörungen|Muenchen|Deutschland
Abstract:
Progressive supranuclear palsy (PSP) is a primary tau-pathy characterized by
neuronal and glial tau-immunoreactive inclusions in different brain regions,
including pons, pallidum, subthalamus and substantia nigra. Vertical gaze palsy
is the hallmark of clinical symptoms, which is later accompanied by horizontal eye
movement deficits ending up in a complete gaze palsy. The vertical premotor
saccadic burst neurons in the rostral interstitial nucleus of the medial longitudinal
fascicle (RIMLF), a major target of tau-pathology in PSP and the oculomotor
nucleus (nIII), were shown to be associated with perineuronal nets (Horn et al.
2008 J. Comp. Neurol. 455:341-352).
Since both areas are specifically spared from tau pathology in Alzheimer\'s
disease a protective role of perineuronal nets (PN) is suggested (Morawski et al.
2010 Neuroscience 169:1347-1363).
In two PSP cases with a complete gaze palsy neighbouring 5µm paraffin sections
containing the RIMLF or nIII were immunostained for hyperphosphorylated tau
protein (AT8) and aggrecan (ACAN), respectively. The overview inspection
revealed rim-like ACAN and AT8-staining in the RIMLF and nIII in both cases.
The close analysis of single neurons revealed that rim-like ACAN-based PNs
were mainly found around AT8-negative neurons. Few AT8-positive neurons
were present with weaker ACAN staining - appearing as interrupted contours.
In conclusion we showed that the presence of PNs within certain brainstem nuclei
may not protect the whole nucleus from degeneration as suggested from
Alzheimer cases, but on a cellular basis the integrity of PNs is affected by taupathology.
BMBF (IFB-01EO0901, Brain-Net-01GI0505)
Kategorie: Poster
Poster 101
Rubrik: 8. Neuroregeneration/Neurodegeneration
Abstract Nr.:
Titel: Differences in spinal cord inflammation induced by the mbp-plp fusion
protein mp4 and mog peptide 35-55 in c57bl/6 mice
Autoren: Stormanns E.(1),Addicks K.(1),Kuerten S.(1),
Adressen:(1)Köln|Institut I für Anatomie|Köln|Deutschland;
email:stefanie.kuerten@uk-koeln.de
Abstract:
Multiple sclerosis (MS) is considered to be an autoimmune disorder of the CNS. It
still remains unclear, however, to what extent particular CNS antigens can cause
specific histopathological differences. Here we demonstrate that experimental
autoimmune encephalomyelitis (EAE) induced by the myelin basic protein (MBP)proteolipid protein (PLP) fusion protein MP4 in C57BL/6 mice shows different
characteristic features in spinal cord histopathology compared to a disease
elicited by MOG peptide 35-55. Mice were immunized with MOG:35-55 and MP4
and were scored daily according to the standard EAE scale over a period of six
months. The degree of spinal cord inflammation was assessed on methylene
blue-stained semi-thin sections in acute EAE and three versus six months after
disease onset, respectively, focusing on the anterolateral, posterior and pyramidal
tract of the lumbar spinal cord. Our results indicate that in both models spinal cord
inflammation was time-dependent being most prominent in acute EAE and
decreasing as disease progressed to the chronic stage. However, differences
pertained to the patterns of inflammation including differences in the extent of
edema formation and the targeting of different fiber tracts. In addition, while MP4induced EAE showed a correlation between the degree of spinal cord
inflammation and the clinical disease severity, such a correlation was absent in
the MOG:35-55 model. The data imply that different patterns of CNS inflammation
can be induced by the use of different CNS antigens in EAE models. These
findings may facilitate the comprehension of the complexity of MS histopathology.
Kategorie: Poster
Poster 102
Rubrik: 8. Neuroregeneration/Neurodegeneration
Abstract Nr.:
Titel: Changes in the expression of genes associated with synaptic plasticity in
the PAHenu2 mouse
Autoren: Schlegel G.(1),Horling K.(2),Santer R.(3),Ullrich K.(4),Schumacher
U.(5),Rune G.(1),
Adressen:(1)University Medical Center Hamburg-Eppendorf|Department of
Neuroanatomy|Hamburg|Germany; email:gschlegel05@googlemail.com;
(2)University Medical Center Hamburg-Eppendorf,|Department of Anatomy and
Experimental Morphology|Hamburg|Germany; (3)University Medical Center
Hamburg-Eppendorf|Department of Pediatrics|Hamburg|Germany; (4)University
Medical Center Hamburg-Eppendorf, Hamburg, Germany|Department of
Pediatrics|Hamburg|Germany; (5)University Medical Center HamburgEppendorf|Department of Anatomy and Experimental
Morphology|Hamburg|Germany
Abstract:
Phenylketonuria (PKU) is a genetic metabolic disorder, which is associated with
severe mental retardation, brain damage and seizures in untreated patients. In
this disorder, mutation of hepatic enzyme Phenylalanine hydroxylase (PAH),
normally converting phenylalanine into tyrosine, results in abundant amounts of
phenylalanine in the blood. The molecular mechanisms by which elevated levels
of phenylalanine cause mental retardation are not known so far. In previous
experiments, we found that treatment of hippocampal slice cultures with high
doses of phenylalanine induced structural changes on hippocampal synapses.
Similar changes were found in the PAH knock out mouse (PAHenu2). To assess
the underlying molecular changes of altered structural synaptic plasticity we used
the PCR-Assay Mouse Synaptic Plasticity RT² ProfilerTM (Qiagen), containing
more than 80 genes associated with synaptic plasticity, as a screening method.
First results show an altered regulation of the following genes: Adcy8, Egr2,
Grm8, Grin2c, Ppp1r14a, Pim1, Ntf5 and CnR1.
Kategorie: Poster
Poster 103
Rubrik: 8. Neuroregeneration/Neurodegeneration
Abstract Nr.:
Titel: Neurological and ophthalmological manifestations of orbital trauma
Autoren: POP E.(1),FOLESCU R.(2),MOTOC A.(1),PETRESCU C.(3),SISU A.(4),
Adressen:(1)UNIVERSITY OF MEDICINE AND PHARMACY "VICTOR BABES"
TIMISOARA|DEPARTMENT OF ANATOMY AND
EMBRYOLOGY|TIMISOARA|ROMANIA; email:alexandra_2987@yahoo.com;
(2)UNIVERSITY OF MEDICINE AND PHARMACY " VICTOR BABES"
TIMISOARA|DEPARTMENT OF ANATOMY AND
EMBRYOLOGY|TIMISOARA|ROMANIA; (3)UNIVERSITY OF MEDICINE AND
PHARMACY "VICTOR BABES"|DEPARTMENT OF ANATOMY AND
EMBRYOLOGY|TIMISOARA|ROMANIA; (4)UNIVERSITY OF MEDICINE AND
PHARMACY"VICTOR BABES"|DEPARTMENT OF ANATOMY AND
EMBRYOLOGY|TIMISOARA|ROMANIA
Abstract:
Early diagnosis and appropriate treatment are necessary because the central
nervous system (CNS) traumatic injuries may be irreversible without early
therapeutic intervention. More than a third of patients with maxillofacial injuries
were contusions and eye injuries, 3% of these produce blindness. More than 90%
of severe injuries result from centrofacial, supraorbital trauma, or frontal sinus
fractures. Four percent of these patients had lesions of the optic nerve, and 23%
paralysis of cranial nerves. If there are deficiencies in both optic nerves
associated deficiency may not be evident by visual stimulation test by successive
light because there is no difference between the two eyes. If pupils are unequal
and not traumatized iris is probably a motor deficiency. CNS complications are:
carotid-cavernous fistula, subdural hematoma, subarachnoid hemorrhage,
meningitis, pneumoencephal, intracranial hypertension, superior orbital fissure
and cavernous sinus penetration, abscesses, cysts and foreign bodies.
Paracarotid space lesions may cause carotid thrombosis and these patients
carotid angiography is recommended, especially if trauma is suspected
carotidian.5% of patients suffer bone fractures and facial fractures have orbital
roof. Complications are rhinorrhea with cerebrospinal fluid, dural fistula and
leakage of cerebrospinal fluid. Manifested as a serous or bloody, in combination
with a rupture of the dura is followed by meningitis in 8.6 - 41% of cases.
Retrobulbar hematomas usually accompany orbital roof fractures, but they rarely
and temporally affect optic nerve function. Middle cranial fossa penetration
through superior orbital fissure can cause intracranial hemorrhage, and total
ophthalmoplegia.
Keywords: cranial nerve, trauma, fractures.
Kategorie: Poster
Poster 104
Rubrik: 9.Peripheres und vegetatives Nervensystem
Abstract Nr.:9
Titel: First evidence for site-specific gene expression of the reelin system in the
human enteric nervous system
Autoren: Ghorbani P.(1),Egberts J.(2),Becker T.(2),Wedel T.(1),Böttner M.(1),
Adressen:(1)CAU Kiel|Anatomisches Insitut|Kiel|Schleswig-Holstein;
email:p.ghorbani@gmx.de; (2)UKSH Campus Kiel|Chirurgie|Kiel|SchleswigHolstein
Abstract:
Background & aims:
Reelin exerts its signal transduction via VLDLR and Lrp8. In the CNS, reelin
coordinates migration and lamination of neurons and regulates synaptic plasticity.
Reelin dysregulation has been implicated in neurological and psychiatric
disorders pointing to its role in maintenance of neural networks The aim of the
study was to determine the expression pattern and localization of reelin in the
human ENS and to monitor the time course of mRNA expression of reelin and its
receptors in the rat intestine.
Material & methods:
Gene expression of reelin and its receptors were analysed in human colonic
specimens. Tissue was sampled from full-thickness sections, separated intestinal
layers and isolated enteric ganglia harvested by laser microdissection (LMD). Colocalization of reelin with the pan-neuronal marker PGP 9.5 was studied by duallabel-immunocytochemistry. The time course of gene expression of the reelin
system was monitored in an ontogenetic study of rat intestines.
Results:
Reelin, VLDLR, and Lrp8 were expressed in all intestinal layers with highest
levels detectable within the tunica muscularis. Site-specific gene expression of
the reelin system was detected in enteric ganglia harvested by LMD. Myenteric
and submucosal ganglia and nerve fibers were immunoreactive for reelin which
co-localized with PGP9.5. Ontogenetic gene expression analysis revealed highest
expression of the reelin system at early postnatal stages.
Conclusions: Reelin and its receptors are strongly expressed in the human ENS.
Reelin is specifically localized in enteric neurons with highest expression levels
during early postnatal life indicating putative functions in the differentiation and
maintenance of the ENS.
Kategorie: Poster
Poster 105
Rubrik: 9.Peripheres und vegetatives Nervensystem
Abstract Nr.:9
Titel: Fibroblast growth factors (fgfs) as new candidates for regulation of sensory
c-fibers in murine airways
Autoren: Böhmer R.(1),Nandigama R.(1),Wiegand S.(1),Kummer
W.(1),Nassenstein C.(1),
Adressen:(1)Justus-Liebig-Universität|Institut für Anatomie und
Zellbiologie|Giessen|Deutschland;
email:christina.nassenstein@anatomie.med.uni-giessen.de
Abstract:
Bronchopulmonary sensory C-fibers (BSCF) play an important role in transducing
airway inflammation into symptoms of asthma such as bronchospasm, mucus
secretion and urge to cough. The mechanisms by which these sensory neurons
are activated in asthma have not yet been worked out.
Members of the Fibroblast Growth Factor (FGF) family are upregulated in the
bronchi of asthmatics, and are expressed by bronchial epithelial cells and located
in basement membranes. Since many terminals of BSCF are located within or
beneath the airway epithelium, we hypothesize that FGFs contribute to an altered
function in BSCF.
We first investigated if BSCF express FGF receptors (FGFR). Neurons of the
sensory vagal ganglia were isolated after retrograde tracing from the airways, and
single-cell RT-PCR was performed. About 60% of all BSCF expressed FGFR1,
whereas FGFR2-4 were not expressed. Further analysis revealed that FGFR1IIIc,
which shows a higher binding affinity to its ligands compared to FGFR1IIIb, was
the most abundant splice variant. Since this receptor is known to bind several
FGFs, we tested which of the possible candidates were expressed in murine lung
and therefore might be able to influence the function of the BSCF. Systematic
analysis showed that FGF1 and 18 were constitutively expressed in the lung, and
FGF2, 7 and 10 could be found in the majority of the samples. All other tested
FGFs were expressed in lower frequency (FGF3, 5, 9, 17, 20, 21 and 23), or
completely absent (FGF4, 6, 8, 16, 22). In a series of further experiments, we will
test if FGFs can increase the excitability of vagal BSCF via FGFR1IIIc and
therefore contribute to the symptoms observed in asthmatics.
Kategorie: Poster
Poster 106
Rubrik: 9.Peripheres und vegetatives Nervensystem
Abstract Nr.:9
Titel: Expression and localization of the snare protein snap-25 in the human
enteric nervous system and its regulation by gdnf in enteric nerve cell cultures
Autoren: Barrenschee M.(1),Harde J.(2),Egberts J.(3),Becker T.(3),Wedel
T.(2),Böttner M.(2),
Adressen:(1)Christian-Albrechts-Universität Kiel|Anatomisches
Institut|Kiel|Deutschland; email:m.barrenschee@anat.uni-kiel.de; (2)ChristianAlbrechts-Universität zu Kiel|Anatomisches Institut|Kiel|Deutschland;
(3)Universitätsklinikum Schleswig-Holstein|Klinik für Allgemeine Chirurgie und
Thoraxchirurgie|Kiel|Deutschland
Abstract:
Background & aims:
The enteric nervous system (ENS) corresponds to the largest nervous system
outside the CNS and thus has also been termed “little brain in the gut”.
Synaptosomal-associated protein 25 (SNAP-25), a member of the SNARE
complex involved in exocytosis of synaptic vesicles, is known to be expressed by
neurons and neuroendocrine cells in the CNS. Since little data are available
concerning SNAP-25 expression in the human ENS, this study was aimed to gain
more detailed insights into the expression and distribution of SNAP-25 in the
human ENS and its regulation by neurotrophic factors.
Material and methods:
Human colonic specimens were processed for SNAP-25 immunohistochemistry
and colocalization was assessed by dual-labeling with the pan-neuronal marker
PGP 9.5. For site-specific gene expression studies (RT-qPCR), material was
obtained from full-thickness sections, tunica muscularis, mucosa, and isolated
enteric ganglia harvested by laser microdissection (LMD). Moreover, SNAP-25
gene and protein expression were analyzed in enteric nerve cell cultures
stimulated with glial cell line-derived neurotrophic factor (GDNF).
Results:
SNAP-25 immunoreactivity was observed in both submucosal/myenteric plexus
and nerve fibers of colonic muscle layers. SNAP-25 gene expression was
detected in full-thickness sections, mucosa, tunica muscularis and myenteric
ganglia isolated by LMD. GDNF treatment increased both protein and gene
expression of SNAP-25 in cultured enteric neurons.
Conclusion:
The findings provide evidence for the first time that SNAP-25 is physiologically
present in the human ENS involving all intramural nerve plexus layers. The data
may help to evaluate altered SNAP-25 expression in gastrointestinal diseases
associated with enteric neuropathies.
Kategorie: Poster
Poster 107
Rubrik: 9.Peripheres und vegetatives Nervensystem
Abstract Nr.:9
Titel: Impaired balance between inhibitory and excitatory intramuscular nerve
fibres in human chagasic megacolon
Autoren: Jabari S.(1),da Silveira A.(2),Quint K.(3),Neuhuber W.(1),Brehmer A.(1),
Adressen:(1)Friedrich- Alexander- Universität Erlangen- Nürnberg|Institut für
Anatomie 1|Erlangen|Deutschland; email:samir.jabari@anatomie1.med.unierlangen.de; (2)Universidade Federal de Uberlândia|Human Anatomy
Sector|Uberlândia|Brazil; (3)Philipps Universität Marburg|Institut für Chirurgische
Forschung|Marburg|Deutschland
Abstract:
Megacolon, chronic dilation of a colonic segment, is a frequent syndrome of
Cagas disease. It is accompanied by an extensive neuron loss which, as shown
recently, results in a partial, selective survival of nitrergic myenteric neurons.
Here, we focused on the balance of intramuscular excitatory (cholin
acetyltransferase/ChAT- immunoreactive) and inhibitory (neuronal nitric oxide
synthase/NOS- as well as Vasoactive Intestinal Peptide/VIP- immunoreactive)
nerve fibres.
From surgically removed megacolonic segments of seven patients, three sets of
kryosections (from non-dilated oral, megacolonic and non-dilated anal parts) were
immunhistochemically triple-stained for ChAT, NOS and VIP. Field
measurements of nerve fibre profiles within the muscle layers were compared
with those of seven age- and region-matched, non-chagasic control patients.
Generally, the nerve fiber density (nerve profile area per muscle layer area) was
significantly reduced in all three chagasic segments as compared to controls. The
relation of NOS- to ChAT-positive fiber area (control about 2 to 1) was changed
towards an overbalance of NOS-fibres: oral part 3 to 1, dilated segment 5 to 1,
anal segment 6 to 1. The portion of VIP/NOS-coreactive nerve fibres increased,
from 2 % (control) to 6 % (dilated and anal chagasic segments).
We suggest that overbalance of inhibitory NOS- and NOS/VIP-fibres related to
excitatory ChAT-fibres may be one factor explaining the massive, long lasting and
irreversible dilation of a colonic segment. Since the overbalance is most
pronounced in the non-dilated anal segment, other components of the motor
apparatus (musculature, interstitial cells of Cajal, submucosal neurons) have to
be considered.
Kategorie: Poster
Poster 108
Rubrik: 9.Peripheres und vegetatives Nervensystem
Abstract Nr.:9
Titel: Gdnf promotes synaptic plasticity in cultured myenteric neurons as revealed
by expression of synaptic vesicle markers
Autoren: Harde J.(1),Barrenschee M.(1),Wedel T.(1),Egberts J.(2),Becker
T.(2),Böttner M.(1),
Adressen:(1)Christian-Albrechts-Universität Kiel|Anatomisches
Institut|Kiel|Deutschland; email:j.harde@anat.uni-kiel.de; (2)Universitätsklinikum
Schleswig-Holstein Campus Kiel|Klinik für Allgemeine Chirurgie und
Thoraxchirurgie|Kiel|Deutschland
Abstract:
Background and aims:
Glial cell line-derived neurotrophic factor (GDNF) is essential for the development
and functional maintenance of the enteric nervous system (ENS). Intestinal
motility is mediated by enteric neurotransmission requiring an intact synaptic
vesicle apparatus. In the present study we investigated markers of synaptic
plasticity in the human ENS and their regulation by GDNF in cultured myenteric
neurons.
Material and methods:
Full-thickness specimens of human colon were screened immunohistochemically
for synaptophysin. Effects of GDNF on synaptophysin and the SNARE protein
synaptobrevin were monitored in enteric nerve cell cultures isolated from rat small
intestine and cultured in defined medium +/- GDNF for 3 weeks. mRNA
expression levels for synaptophysin and synaptobrevin were assessed by
quantitative RT-PCR. Additionally, colocalization of synaptic vesicle markers with
the pan-neuronal marker PGP 9.5 was assessed.
Results:
Human colonic specimens showed immunoreactivity of synaptophysin in both
myenteric and submucosal ganglia as well as in nerve fibers. GDNF treatment
promoted synaptic plasticity in cultured myenteric neurons as demonstrated by
the dose-dependent increase of immunoreactive signals and mRNA expression
levels of synaptophysin and synaptobrevin. Both synaptic vesicle markers colocalized with PGP 9.5 and were organized in granular accumulations.
Conclusions:
The increased expression of synaptophysin and synaptobrevin at both mRNA and
protein levels after GDNF treatment suggests that GDNF enhances synaptic
plasticity of the ENS.
The granular accumulations most likely represent nerve fiber varicosities
resembling sites of enteric neurotransmission. Both synaptic vesicle markers are
promising candidates to further characterize enteric neuropathies associated with
human intestinal motility disorders.
Kategorie: Poster
Poster 109
Rubrik: 9.Peripheres und vegetatives Nervensystem
Abstract Nr.:9
Titel: Expression and regulation of the adrenomedullin signaling system in murine
principal sympathetic neurons
Autoren: Wessels L.(1),Pfeil U.(1),Faulhammer P.(1),Weißmann N.(2),Kummer
W.(1),
Adressen:(1)Justus-Liebig-University Giessen|Institut für Anatomie und
Zellbiologie|Gießen|DE; email:larswessels@gmx.de; (2)Justus-Liebig-University
Giessen|Excellence Cluster Cardio-Pulmonary System, Chair for Molecular
Mechanisms of Emphysema, Hypoxia and Lung Aging|Gießen|DE
Abstract:
Peptides of the adrenomedullin (AM) family (AM, intermedin = AM2, calcitonin
gene-related peptide = CGRP) induce facilitation of synaptic transmission. Here,
we investigated regulation of this signalling system in the murine superior cervical
ganglion (SCG) by q-PCR, qualitative single cell RT-PCR, in situ-hybridisation,
and immunohistochemistry. In the whole ganglion, baseline expression of AM
exceeds that of AM2 by a factor of 23, and all components of the receptor system
(calcitonin receptor-like receptor = CLR, receptor activity modifying proteins1-3 =
RAMP1-3)
are
expressed.
Qualitative
single
cell
RT-PCR
and
immunohistochemistry identified principal sympathetic neurons as a source of
AM. In freshly explanted SCGs, 6 h incubation with either muscarine or CGRP
strongly upregulated AM-mRNA while nicotine was less effective. These effects
were sensitive to specific receptor blockers (atropine, CGRP8-37, and
mecamylamine, respectively). Exposure of C57Bl6N mice to hypoxia (10% O2)
for 24 hours resulted in about 3-fold AM-, AM2- and RAMP1-mRNA upregulation
which was significantly attenuated both by surgical decentralization and sham
operation 7 days prior to hypoxia exposure. In conclusion, expression of AM by
principal sympathetic neurons is upregulated by metabotropic receptors for
mediators contained within intraganglionic terminals (acetylcholine, CGRP) and
after housing the animals in hypoxic atmosphere. This effects may be partly but
not entirely driven by preganglionic activity. Due to the facilitating effect on
neuronal excitability of AM/AM2 this pathway might be involved in adaptation of
sympathetic neurons in settings of enhanced cardiovascular activity.
Kategorie: Poster
Poster 110
Rubrik: 9. Peripheres und vegetatives Nervensystem
Abstract Nr.:
Titel: Gabaergic innervation of the ciliary ganglion in monkey
Autoren: Barnerssoi M.(1),Lienbacher K.(1),Zeeh C.(1),May P.(2),Mustari
M.(3),Horn A.(1),
Adressen:(1)LMU Muenchen|Anatomisches Institut, Lehrstuhl
I|Muenchen|Deutschland; (2)University of Mississippi Medical Center|Department
of Neurobiology and Anatomical Sciences|Jackson|USA; (3)Washington National
Primate Research Center, University of Washington|Department of
Ophthalmology|Seattle|USA
Abstract:
The vertebrate ciliary ganglion (CG) is situated behind the ocular bulb between
the optic nerve and lateral rectus muscle. It controls lens accommodation (ciliary
muscle) and pupillary constriction (iris sphincter muscle). Currently, only the
preganglionic, cholinergic neurons of the Edinger-Westphal nucleus (EWpg) are
considered to supply afferent nerve endings to this ganglion. Avian studies
indicated that some postganglionic CG neurons may also be controlled by GABA,
but its source is unknown.
Here we investigated the CG of the macaque monkey for the presence of
GABAergic terminals. Immunostaining for the GABA-synthesizing enzyme,
glutamate decarboxylase (GAD), revealed a population of ganglionic neurons with
a dense supply of GAD-positive terminals. These cells make up 17,5% of the total
CG neuron population, and they are evenly distributed throughout the ganglion.
To identify the source of the GABAergic innervation, the CG of two monkeys, that
had received a tracer injection (biotinylated dextran-amine; BDA) into the
oculomotor nucleus including the overlying EWpg and supraoculomotor area
(SOA) were analysed for the presence of tracer-labelled GAD-positive terminals
with double immunofluorescence methods. Both cases revealed tracer-labelled
afferent terminals in the CG. However, only the more lateral of the two injections
resulted in BDA-positive terminals that were double labeled for GAD. These
results indicate that GABAergic inputs to CG neurons are also present in a
primate, and that these inputs most likely arise from neurons located in SOA
lateral to EWpg. The function of this GABAergic innervation of the CG remains to
be clarified.
DFG (HO 1639/4-3), NIH EY014263, EY020744, RR000166
Kategorie: Poster
Poster 111
Rubrik: 9. Peripheres und vegetatives Nervensystem
Abstract Nr.:
Titel: Gpr91 and gpr99 in murine sympathetic and sensory neurons
Autoren: Paddenberg R.(1),Diehl J.(2),Gries B.(2),Pfeil U.(2),Raju R.(2),Kummer
W.(2),
Adressen:(1)Justus-Liebig Universität|Institut für Anatomie und
Zellbiologie|Gießen|Deutschland; email:Renate.Paddenberg@anatomie.med.unigiessen.de; (2)Justus-Liebig-Universität|Institut für Anatomie und
Zellbiologie|Gießen|Deutschland
Abstract:
GPR91 and GPR99 are G-protein-coupled receptors which are activated by the
citric acid cycle intermediates succinate and alpha-ketoglutarate, respectively. In
the kidney, succinate binding to GPR91 stimulates the release of renin. GPR99
mRNA is highly expressed in kidney and also detectable in testis. Aim of our
study was to analyse expression and function of GPR91 and GPR99 in neuronal
cells.
GPR91 and GPR99 mRNA were detected by RT-PCR in sympathetic (superior
cervical ganglia, stellate ganglia) and sensory ganglia (trigeminal ganglia, DRG).
In western blots of DRG and trigeminal ganglia we obtained a single band of
appropriate molecular mass (37 kDa) and the signal was not detectable after
preabsorption
of
the
antibody
with
the
corresponding
peptide.
Immunohistochemistry for GPR91 and GPR99 revealed ubiquitous expression of
both receptors in all neurons of sympathetic and sensory ganglia. In cultured
DRG neurons immunohistochemically labeled for PGP 9.5 or NF-200 neurite
morphology was analysed using the IMARIS software. Application of succinate or
alpha-ketoglutarate significantly affects the neurite organisation: Neurite length,
numbers of branch points, segments and terminal points were increased as
compared to untreated controls. Since plasma concentrations of succinate and
alpha-ketoglutarate are distinctly increased in patients suffering from diabetic
ketoacidose GPR91 and GPR99 may participate in the development of sensory
neuropathy, a fatal complication in diabetes.
Kategorie: Poster
Poster 112
Rubrik: 10.Zentrales Nervensystem/Signaltransduktion und Verschaltung
Abstract Nr.:1
Titel: An in vitro model for scar formation to study the mechanisms of scarreducing treatments used in spinal cord injury
Autoren: Vogelaar C.(1),Krafft S.(2),Ziegler B.(2),Müller H.(3),
Adressen:(1)Johannes Gutenberg-Universität Mainz|Institut für Mikroskopische
Anatomie und Neurobiologie|Mainz|Deutschland;
email:tineke.vogelaar@unimedizin-mainz.de; (2)Heinrich-Heine Universität
Düsseldorf|Labor für molekulare Neurobiologie, MNR
Klinik|Düsseldorf|Deutschland; (3)Heinrich-Heine Universität Düsseldorf|Labor für
molekulare Neurobiologie|Düsseldorf|Deutschland
Abstract:
Spinal cord injury leads to permanent damage of axon tracts and impairment of
sensory and motor functions. Lesion-induced fibrous scarring is a major
impediment for regeneration of injured axons in the CNS. The collagen-rich scar
contains axon growth inhibitory factors. In our laboratory a pharmacological
treatment was developed, transiently suppressing fibrous scarring (Klapka et al,
2005). This “anti-scarring treatment” (AST) consists of local application of an iron
chelator and cyclic AMP, inhibiting collagen synthesis by invading fibroblasts.
AST treatment stimulated regeneration of various axon tracts, leading to
improvements in locomotor recovery.
In order to study the molecular mechanisms of AST we used an in vitro model for
scar formation. In this model, fibroblasts and astrocytes in co-culture form scarlike clusters after addition of TGF-beta1. We investigated in more detail the
mechanisms of scar formation. Incubation with BrdU revealed that clusters are
formed by proliferation. Live cell imaging showed that cluster formation also
involved reorganization of the existing fibroblast layer and scar-like contraction of
the cluster. Migration of cells into the cluster was rarely seen.
Immunohistochemical analysis showed that mature clusters contain various
growth inhibitory proteins, like CSPGs, sema-3A, and tenascin-C. Cultivation of
cortical neurons on these cluster revealed that axon growth was reduced. Axons
tended to stay close to astrocytes and did not cross the barriers of inhibitory
molecules that are usually around the clusters. We will use the model to study
and improve existing treatments, but also to find new treatment strategies.
Kategorie: Poster
Poster 113
Rubrik: 10.Zentrales Nervensystem/Signaltransduktion und Verschaltung
Abstract Nr.:1
Titel: Characterization of mice lacking the autism spectrum disorders associated
protein prosap2
Autoren: Janssen A.(1),Schmeisser M.(1),Böckers T.(2),Bockmann J.(1),
Adressen:(1)Ulm|Anatomie und Zellbiologie|Ulm|Deutschland; email:annalena.janssen@uni-ulm.de; (2)Ulm|Anatomie und Zellbiologie|U|Deutschland
Abstract:
Autism spectrum disorders (ASD) include impairments in social interaction,
communication difficulties and repetitive behaviour as symptoms. Etiology of this
neurodevelopmental disorder is still unknown but it is assumed that disturbed
development or misfunction of glutamatergic synapses plays a crucial role. Cases
of ASD are linked to mutations in the ProSAP2 gene, a member of a family of
scaffold proteins that are essential for development and molecular organization of
glutamatergic synapses. The mutations lead to truncated forms of ProSAP2 and
probably affect its function and localization to postsynaptic densities (PSD). To
study the role of ProSAP2 in the etiology of ASD, we created a ProSAP2 knockout (KO) mouse. mRNA analysis verified that deletion of the targeted exon leads
to stop of translation in the subsequent exon, resulting in a truncated protein
missing important domains that mediate protein-protein interaction and targeting
of ProSAP2 to PSDs. Inexplicably, on Western Blots, where ProSAP2 is typically
reflected as a triplet band, only two out of three bands are absent in mutant mice.
Whether the remaining band - also seen in all ProSAP2 KO mouse models of
other groups - indeed is a form of ProSAP2 is unclear. Irregardless of this,
analysis of protein composition of PSDs uncovered diminished expression levels
of glutamate receptor subunits and various scaffold proteins in KOs. Interestingly,
ProSAP1 is up-regulated in ProSAP2 mutants, probably reflecting a
compensatory mechanism. In summary, our data support the assumption that
misfunction of ProSAP2 leads to disturbed organization of glutamatergic
synapses which might then result in development of ASD.
Kategorie: Poster
Poster 114
Rubrik: 11.Neuroimmunologie
Abstract Nr.:1
Titel: Endogenous tgf-beta signalling in primary microglia - evidence for autocrine
inhibition of pro-inflammatory microglia phenotype in vitro
Autoren: Spittau B.(1),Wullkopf L.(1),Zhou X.(1),Krieglstein K.(1),
Adressen:(1)Albert-Ludwigs-Universität Freiburg|Institut fur Anatomie und
Zellbiologie, Abteilung für Molekulare Embryologie|Freiburg|Deutschland;
email:bjoern.spittau@anat.uni-freiburg.de
Abstract:
Transforming growth factor beta 1 (TGF-beta1) plays important roles in a variety
of physiological and pathophysiological processes in the central nervous system.
TGF-beta1 deficient mice show progressive neurodegeneration within the first
postnatal weeks accompanied by strong microgliosis. Moreover, extensive
activation of microglia in TGF-beta1 mutants has further been shown in otherwise
uninjured brain areas, indicating an essential function of TGF-beta1 as a potent
endogenous regulator of microglia activity. Here we analysed the expression of
TGF-beta isoforms in primary microglia and addressed the role of endogenous
TGF-beta signalling for the regulation of microglia activity in vitro. We
demonstrate that microglia express and secrete high levels of TGF-beta1 under
unstimulated conditions. Further, treatment with recombinant TGF-beta1 resulted
in Smad2 phosphorylation and nuclear accumulation indicating an active TGFbeta signalling pathway in primary microglia. Tieg1/Klf10, a well described TGFbeta target gene, was rapidly upregulated in primary microglia. Using microgliaconditioned medium we demonstrate that microglia-derived TGF-beta1 is able to
activate the TGF-beta signalling pathway in microglia cultures in an autocrine
manner. Moreover, blocking endogenous TGF-beta signalling using a TGF-beta
receptor type I inhibitor resulted in increased expression of the classical microglia
activation markers iNOS and IL6 and downregulation of the alternative activation
marker Arg1. Together, these data strongly support the role of endogenous TGFbeta1 in regulating microglia activity to prevent extensive activation and microgliamediated neurotoxicity.
Kategorie: Poster
Poster 115
Rubrik: 11.Neuroimmunologie
Abstract Nr.:
Titel: Cytokine expression in the mouse brain microvascular endothelial cell line
cEND after dexamethasone treatment
Autoren: Burek M.(1),Förster C.(1),
Adressen:(1)Universität Würzburg|Klinik und Poliklinik für
Anästhesiologie|Würzburg|Deutschland; email:Burek_M@klinik.uni-wuerzburg.de
Abstract:
Multiple sclerosis (MS) is an autoimmune disease of the central nervous system
(CNS). It is characterized by the immune cell migration into the CNS,
demyelination and various degrees of axonal damage. Compromised blood-brain
barrier (BBB) integrity is a major hallmark of active MS. Migration of immune cells
into CNS is largely regulated by cytokines and the targeting of this process is one
of the therapeutic strategies of MS. Brain endothelial cells express distinct
cytokines along with their receptors. Here, we characterized the expression
pattern of selected cytokines and their receptors in the murine in vitro BBB model,
cEND. Glucocorticoids are used in the MS therapy to treat the acute relapses of
the disease. Thus, we also examined the cytokine expression in cEND after
treatment with dexamethasone. For this purpose, we conducted 96-well real-time
transcription polymerase chain reaction (qRT-PCR) arrays to quantitatively
compare the mRNA expression. We detected basal mRNA expression of different
classes of cytokines and their receptors. Moreover, significant changes in mRNA
expression of Csf3, Ccl12, Cxcr3, Il17f and Kit were detected after
dexamethasone treatment.
We confirmed the results of qRT-PCR array in qPCR and additionally examined
the expression changes of some cytokines after treatment of cEND with MS
patient sera with or without addition of dexamethasone. In conclusion, cEND cell
line can be used to explore the changes in BBB function under proinflammatory
conditions, as well as to study the influence of endothelial cytokines on the other
components of the neurovascular unit.
Kategorie: Poster