C.V.:
Diefenbach, T. J.
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Curriculum Vitae
PART I: General Information
DATE PREPARED:
February 16, 2016
Name:
Thomas John Diefenbach, Ph.D.
Office Address:
Department of Neurology
Children’s Hospital Boston
320 Longwood Avenue, EN282
Boston MA 02115
Home Address:
67 Willow Avenue #1
Somerville, MA 02144
Work E:Mail:
Thomas.Diefenbach@childrens.harvard.edu
Work FAX:
617-730-0242
Place of Birth:
Edmonton, Alberta, Canada
Education:
1990
B.Sc. (Specialization) in Zoology
University of Alberta, Edmonton, Alberta, Canada
1995
Ph. D. in Biological Sciences
University of Alberta, Edmonton, Alberta, Canada
Postdoctoral Training:
1994
Department of Biological Sciences Laser Scanning Confocal
Microscopy Workshop (Certificate)
Instructor: Dr. R. Bhatnager
Electron Microscope Lab
Department of Biological Sciences
University of Alberta, Edmonton, Alberta, Canada
1995
Department of Biological Sciences Fura-2 Calcium Imaging
Workshop
Instructor: Dr. Peter B. Guthrie
Department of Anatomy and Neurobiology
Colorado State University
Fort Collins, CO
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1995-1999
Postdoctoral Fellow
Laboratory of Dr. Stanley B. Kater
Department of Neurobiology and Anatomy
University of Utah School of Medicine
Salt Lake City, Utah
1999-2002
Postdoctoral Fellow
Laboratory of Dr. Daniel G. Jay
Department of Physiology
Tufts University School of Medicine
Boston, Massachusetts
Diefenbach, T. J.
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Hospital or Affiliated Institution Appointments:
2002-2005
Research Associate
Laboratory of Dr. Daniel G. Jay
Department of Physiology
Tufts University School of Medicine
Boston, Massachusetts
2005-present Instructor of Neurology
Harvard Medical School
Department of Neurology
Children’s Hospital, Boston MA
Manager of Imaging Operations
Developmental Disabilities Research Center Imaging Core
Department of Neurology
Children’s Hospital, Boston MA
Visiting Scientist
Department of Physiology
Tufts University School of Medicine
Boston, Massachusetts
Major Administrative Responsibilities:
1990-1994
Graduate Student Association Representative
Department of Zoology and Biological Sciences
University of Alberta
1992-1993
Graduate Student Representative
Department of Zoology Council
University of Alberta
2001-2002
Postdoctoral Fellow Representative
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Diefenbach, T. J.
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Department of Physiology Faculty Council
Tufts University School of Medicine
Professional Societies:
1993-present The Society for Neuroscience
2000-present The American Society for Cell Biology
Awards and Honors:
1985
Alexander Rutherford Scholarship for Undergraduate Study
Government of Alberta
1990-1992
Graduate Teaching Assistantship
Department of Zoology
University of Alberta
1992-1995
Studentship
Alberta Heritage Foundation for Medical Research
1995
Graduate Teaching Assistantship
Department of Biological Sciences
University of Alberta
1995-1998
Postdoctoral Fellowship
Alberta Heritage Foundation for Medical Research
Part II: Research, Teaching, and Clinical Contributions
A. Narrative report
My research career began as research technician in the laboratory of Dr. William C.
Mackay at the Department of Zoology, University of Alberta, 1986-87. There I
performed laboratory and field work in fish environmental physiology, undertook data
analysis on the mainframe computer, and produced technical and graphic artwork for
publication. In 1988 I conducted an independent study with Dr. Jeffrey I. Goldberg on a
project entitled “Using intracellular microelectrodes, electrophysiologically identify and
characterize neurons involved in the feeding motor circuit of Helisoma trivolvis.” From
1988-1990, as research assistant in the laboratory of Dr. Jeffrey I. Goldberg, Department
of Zoology, University of Alberta, I worked on “Immunohistochemical characterization
of the postembryonic development of serotonergic neurons in wildtype and inbred strains
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of the ramshorn snail, Helisoma trivolvis.” I again undertook independent study from
1989-1990, this time with Dr. Sudarshan K. Malhotra of Department of Zoology,
University of Alberta, on the project “Immunohistochemical characterization of
intermediate filament proteins in astroglia of wildtype and dystrophic hamster CNS”, in
addition to performed transmission electron microscopy of cerebellar neurons. Also from
1989-1990, I served as research assistant in the laboratory of Dr. Andrew N. Spencer,
Department of Zoology, University of Alberta, on a project entitled “Using whole cell
voltage and current clamp, characterized whole cell ion currents of sensory neurons in
vitro of the ocelli of the jellyfish, Polyorchis panicillatus.” I also performed
immunocytochemistry and immunogold transmission electron microscopy of cultured
neurons. I performed my doctoral thesis research, 1990-1995, under the supervision of
Dr. Jeffrey I. Goldberg, Department of Biological Sciences, University of Alberta. Thesis
title: Development of an Identified Serotonergic Neuron in Embryos of Helisoma
trivolvis: Developmental and Behavioral Consequences of Serotonin Expression. This
study involved extensive examination of neurite outgrowth in situ. For this work I
utilized the techniques of laser scanning confocal microscopy, time lapse video
microscopy, differential interference contrast microscopy, high performance liquid
chromatography, immunohistochemistry, and intracellular microelectrode recording of
electrical activity of neurons in vivo.
I was then awarded the position of Postdoctoral Fellow in the laboratory of Dr. Stanley B.
Kater, Department of Neurobiology and Anatomy, University of Utah School of
Medicine, Salt Lake City, Utah, 1995-1999. During this period I undertook several
specific studies: (1) membrane turnover in neuronal growth cones from ciliary ganglion
of chick embryos; (2) history-dependent responsiveness to laminin-coated microspheres
using optical tweezers and electrical stimulation in growth cones of chick dorsal root
ganglion neurons; and (3) substrate-dependent effects of electrical stimulation on
intracellular calcium responses employing single cell FURA-2 ratiometric calcium
imaging.
Subsequently, as Postdoctoral Fellow in the laboratory of Dr. Daniel G. Jay, Department
of Physiology, Tufts University School of Medicine, Boston, Massachusetts, 1999-2002,
I undertook the study of myosins 1c, IIA, IIB in growth cone motility of chick DRG and
ciliary ganglion neurons using a protein knock-down strategy, chromophore-assisted laser
inactivation (CALI). I also performed in vivo imaging of growth cones in the retinotectal
projection of zebrafish and deconvolution analysis of the neuronal cytoskeleton.
Furthermore, I was responsible for maintenance of microscope/computer/laser equipment
in the laboratory.
During the period 2002-2005, as research associate in the laboratory of Dr. Daniel G. Jay,
Department of Physiology, Tufts University School of Medicine, Boston, Massachusetts,
I continued my study of myosins in growth cone motility with a focus on myosins Va and
VI in the trafficking of membraneous endosomes in neuronal growth cones of cultured
chick ciliary ganglion neurons utilizing a protein knock-down strategy, chromophoreassisted laser inactivation (CALI). I also undertook the imaging of membrane trafficking
in growth cones using the membrane dye, FM1-43 as well as the configuration of a
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custom total internal reflection fluorescence microscopy (TIRFM) optical platform for
imaging of GFP-labeled myosins in living growth cones. I participated in collaborative
work to develop GFP-myosin constructs for live-cell imaging studies and also conducted
fluorescence speckle microscopy of actin filament dynamics in living growth cones.
During this period, I developed a novel method for expression of fusion proteins in
postmitotic cells, for which I am presently pursing a patent application in collaboration
with Tufts University.
Currently, as Manager of Imaging Operations of the Developmental Disabilities Research
Center (DDRC) Imaging Core in the Department of Neurology, Children’s Hospital,
Boston under the supervision of Dr. Scott Pomeroy, Chairman of Neurology, I manage
and maintain the confocal microscopy imaging facility along with three other imaging
systems, train users in microscopic techniques, and collaborate with Neurology faculty on
various research projects. From August to September, 2005, I was responsible for the
design of a new imaging core facility and consolidation and relocation of our imaging
systems in collaboration with CHB Facilities Management and their architects.
My teaching experience includes the private tutoring of over 100 students in grades 7 to
12 mathematics, chemistry and biology, and volunteer tutoring of disadvantaged children,
at Hardesty Junior High School, Edmonton, Alberta (1983-1995). In addition, I taught
the laboratory component of Zoology 342 (Animal Physiology, 6 semesters 1990-1992; 2
semesters, 1995) for the Department of Biological Sciences, University of Alberta. From
1990-1995, I undertook the instruction and supervision of undergraduate students and
technicians for the Department of Biological Sciences, University of Alberta.
Since 1995, I have presented my research findings via seminars at regional, national, and
international venues, and through poster and oral presentations to the Canadian Society of
Zoologists, the IBRO World Congress of Neuroscience, and the Society for
Neuroscience, among others. I am first author on nine reports published in peer-reviewed
journals, and I contributed chapters regarding methods of protein analysis to two texts
published in 2005.
D. Report of Teaching
1. Local contributions
a.
Medical School/School of Dental Medicine/Division of Medical
Sciences course for medical/dental/Ph.D. student
1999-2002
2.
Instruction and supervision of rotation students, instruction of
graduate students/department members/visiting scholars.
Department of Physiology, Tufts University School of Medicine
Regional Contributions
Invited Presentations
2001 Seminar Series
Wellman Laboratories of Photomedicine
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Department of Dermatology
Harvard Medical School at Massachusetts General Hospital
Boston, MA
Chromophore-assisted laser inactivation as a tool to investigate
protein function in cells: myosin II function in neurons
2002
Seminar Series
Department of Anatomy and Structural Biology
Albert Einstein School of Medicine
The Bronx, New York City, NY
Functional contributions of distinct myosins to growth cone
motility
National Contributions
Invited Presentation
1999 Seminar Series
Division of Biology Neurobiology Biology Seminars
California Institute of Technology
Pasadena, CA
A role for stimulus history in the responsiveness of the
neuronal growth cone
International Contributions
Invited Presentation
1995 Symposium in Neuronal Regeneration and Development
Proceedings of the 4th International Congress of Comparative
Physiology and Biochemistry
Birmingham, England
Developmental and behavioral roles of serotonin from an
identified neuron in embryos of the freshwater snail, Helisoma
trivolvis
Part III: Bibliography
Research Papers
1.
Diefenbach TJ, Elbrink J, Malhotra SK. Characterization of intermediate
filament proteins in astroglia of hamster cerebellum. Cytobios 1989;59:193-206.
2.
Diefenbach TJ, Goldberg JI. Postembryonic expression of the serotonin
phenotype in Helisoma trivolvis: comparison between laboratory-reared and
wildtype strains. Can J Zoo 1990;68:1382-9.
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Diefenbach, T. J.
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3.
Diefenbach TJ, Koehncke NK, Goldberg JI. Characterization and development
of rotational behavior in Helisoma embryos: role of endogenous serotonin. J
Neurobiol 1991;22:922-34.
4.
Diefenbach TJ, Elbrink J, Malhotra SK. Glial fibrillary acidic protein, J1-31
antigen and vimentin in adult hamster brain: an immunohistochemical study.
Cytobios 1991;65:39-53.
5.
Martel A, Diefenbach TJ. Effects of body size, water current and microhabitat
on mucous-thread drifting in post-metamorphic gastropods Lacuna spp. Marine
Ecol Prog Ser 1993:99:215-20.
6.
Goldberg JI, Koehncke NK, Christopher KJ, Neumann C, Diefenbach TJ.
Pharmacological characterization of a serotonin receptor involved in an early
embryonic behavior of Helisoma trivolvis. J Neurobiol 1994;25(12):1545-57.
7.
Diefenbach TJ, Sloley BD, Goldberg JI. Neurite branch development of an
identified serotonergic neuron from embryonic Helisoma: Evidence for
autoregulation by serotonin. Dev Biol 1995;167(1): 282-93.
8.
Diefenbach TJ, Koss R, Goldberg JI. Early development of an identified
serotonergic neuron in Helisoma trivolvis embryos: serotonin expression, deexpression, and uptake. J Neurobiol 1998;34:361-76.
9.
Kaufman WR, Sloley BD, Tatchell RJ, Zbitnew GL, Diefenbach TJ, Goldberg
JI. Quantification and cellular localization of dopamine in the salivary gland of
the ixodid tick Amblyomma hebraeum. Exp Appl Acarol 1999;23:251-65.
10.
Diefenbach TJ, Guthrie PB, Stier H, Billups B, Kater SB. Membrane recycling
in the neuronal growth cone revealed by FM1-43 labeling. J Neurosci
1999;19:9436-44.
11.
Diefenbach TJ, Guthrie PB, Kater SB. Stimulus history alters behavioral
responses of neuronal growth cones. J Neurosci 2000;20:1484-94.
12.
Lamb RF, Roy C, Diefenbach TJ, Vinters HV, Johnson MW, Jay DG, Hall A.
The TSC1 tumour suppressor hamartin regulates cell adhesion through ERM
proteins and the GTPase Rho. Nat Cell Biol 2000;2(5):281-7.
13.
Diefenbach TJ, Latham VM, Yimlamai D, Liu C, Herman IM, Jay DG. Myosin
1c and myosin IIB serve opposing roles in lamellipodial dynamics of the neuronal
growth cone. J Cell Biol 2002;158:1207-17.
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Diefenbach, T. J.
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14.
Koss R, Diefenbach TJ, Kuang S, Doran SA, Goldberg JI. Coordinated
development of identified serotonergic neurons and their target ciliary cells in
Helisoma trivolvis embryos. J Comp Neurol 2003;457:313-25.
15.
Wang F-S, Liu C-WA, Diefenbach TJ, Jay DG. Modeling the role of myosin 1c
in neuronal growth cone turning. Biophysical J 2003;85:3319-28.
Book Chapters
1.
Diefenbach TJ, Jay DG. Probing protein function in cells using CALI. In: Dunn
MJ, Jorde LB, Little PFR, Subramaniam S, editors. Encyclopedia of Genetics,
Genomics, Proteomics and Bioinformatics. 1st Edition (4 volumes). Indianapolis,
IN: Wiley Publishing, Inc., 2005.
2.
Diefenbach TJ, Jay DG. Methods for protein knockdown using chromophoreassisted laser inactivation in living cells and tissues. In: Celis JE, Carter N,
Simons K, Small JV, Hunter T, Shotton D, editors. Cell Biology-A Laboratory
Handbook. 3rd Edition (4 volumes). New York, NY: Academic Press, Elsevier
Science, 2005.
Abstracts
(* denotes presenter)
1.
Diefenbach TJ*, Goldberg JI. Selective effects of serotonin on rotational and
feeding behaviors of Helisoma trivolvis embryos. Poster presentation. Bull Can
Soc Zool 1990;21:38.
2.
Goldberg JI, Diefenbach TJ*, Koehncke NK. Behavioral and pharmacological
evidence for a new role of identified serotonergic neurons in embryos of
Helisoma trivolvis. Oral presentation. Third IBRO World Congress of
Neuroscience 1991; Abstract # P42.31.
3.
Diefenbach TJ*, Koehncke NK, Goldberg JI. Role of identified serotonergic
neurons in generating rotational behavior in Helisoma embryos. Poster
presentation. Soc Neurosci Abstr 1991;17:549.
4.
Goldberg JI*, Koehncke NK, Price CJ, Diefenbach TJ. Pharmacological
characterization of serotonin receptors in the snail, Helisoma trivolvis. Poster
presentation. Bull Can Soc Zool 1992;23:59.
5.
Goldberg JI*, Diefenbach TJ*. Early development of an identified serotonergic
neuron in embryos of the gastropod, Helisoma trivolvis. Poster presentation. Soc
Neurosci Abstr 1992;18(1-2):602.
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Diefenbach, T. J.
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6.
Diefenbach TJ*, Goldberg JI. Evidence for autoregulation of neurite branch
development in identified serotonergic neurons from Helisoma embryos. Oral
presentation. Soc Neurosci Abstr 1992;18:602.
7.
Diefenbach TJ*, Koehncke NK*, Christopher KJ, Neuman C, Goldberg JI.
Pharmacological characterization of a serotonin receptor from molluscan
embryos. Poster presentation. Soc Neurosci Abstr 1993;19:1166.
8.
Diefenbach TJ*, Goldberg JI. Distinct neurotransmitter phenotypes in
homologous embryonic neurons from Helisoma trivolvis and Lymnaea stagnalis.
Oral presentation. Bull Can Soc Zool 1994;25:45.
9.
Diefenbach TJ*, Goldberg JI. Developmental and behavioral roles of serotonin
from an identified neuron in embryos of the freshwater snail, Helisoma trivolvis.
Oral presentation. Physiol Zool 1995;68:163.
10.
Goldberg JI*, Koss R, Diefenbach TJ. Anatomical evidence of sensory and
motor function in early serotonergic neurons from Helisoma embryos. Poster
presentation. Soc Neurosci Abstr 1996;22:654.
11.
Diefenbach TJ*, Kater SB. Neuronal growth cone responses are determined by
prior experience. Poster presentation. Soc Neurosci Abstr 1997;23:602.
12.
Diefenbach TJ*, Kater SB. Substrate-dependent responses of neuronal growth
cones to collapse-inducing stimulation. Poster presentation. Soc Neurosci Abstr
1998;24:29.
13.
Guthrie PB, Diefenbach TJ, Stier H, Billups B, Kater SB. Membrane recycling in
the neuronal growth cone: Two distinct pools revealed by FM1-43 labeling. Soc
Neurosci Abstr 1999;25:1023.
14.
Diefenbach TJ*, Latham V, Herman IM, Jay DG. Myosin II inactivation
selectively alters growth cone morphology and retrograde f-actin flow. Poster
presentation. Mol Biol Cell 2000;11(Supplement):2820.
15.
Yimlamai D*, Diefenbach TJ, Jay DG. Molecular cloning of the zebrafish
DSCAM homolog. Poster presentation. Soc Neurosci Abstr 2001;27:1818.
16.
Diefenbach TJ*, Latham VM, Yimlamai D, Liu C, Herman IM, Jay DG.
Opposing roles for myosins 1c and IIB in growth cone motility. Poster
presentation Mol Biol Cell 2002;13(Supplement):179a-180a.
17.
Diefenbach TJ*, Nascimento AAC, Espreafico EM, Jay DG. Opposing roles for
myosin V and VI in filopodial extension and endosomal trafficking in the
neuronal growth cone. Slide presentation. Soc Neurosci Abstr 2004; Program No.
257.2.