The Effects Of Nitric Oxide On The
Development Of Alzheimer's Disease
Caroline Hsiao
Need
http://www.alz.org/advocacy/2006program/images/charts/5.gif
Need
http://msnbcmedia1.msn.com/i/msnbc/Components/Art/HEALTH/070319/AP_ALZHEIM
ERS.gif
Plaques
Plaques
Tangles in Neuron
Alzheimer’s Disease
http://pampee.20gig.com/wpcontent/uploads/2009/04/alzheimersdisease.jpg
http://www.alzheimer.ca/english/disease/
whatisit-intro.htm
Alzheimer’s Disease
http://www.topnews.in/health/files/alzheimers-brain.jpg
Nitric Oxide As A Free Radical
http://www.inchem.org/documents/jecfa/jec
mono/40abcj11.gif
http://www.inchem.org/documents/jecfa/jec
mono/40abcj11.gif
Oxidative Stress
• Cigarette smoke is used as an example shown here
http://healthy-lifestyle.most-effective-solution.com/wpcontent/uploads/2007/10/oxidative-stress.jpg
Age-Dependent Neurodegeneration and AlzheimerAmyloid Plaque Formation in Transgenic Drosophila
Greeve, and
Kretzschmar,
Investigation of Free Radicals in Cigarette
Mainstream Smoke
M. Ghosh and P. Ionita
Dissecting the pathological effects of human
A40 and A42 in Drosophila: A potential model
for Alzheimer’s disease
By: Iijima and Liu
Neuroserpin Binds A and Is a Neuroprotective
Component of Amyloid Plaques in
Alzheimer Disease
By: Kinghorn and Crowther
Purpose
The purpose of this experiment was to find
if preservatives have any effect on the
development of Alzheimer's disease.
Hypothesis
Preservatives will cause an increase in
the development of Alzheimer's
symptoms
Experimental Design
Protocols Used:
Gritta Tettweiler, Mathieu Miron, Mark
Jenkins, 2005
ELIZABETH J. HARRY, KIT POGLIANO,
AND RICHARD LOSICK 1995
The Effects of Nicotine and Nitric Oxide on the development of Amyloid beta plaques
in wild type and APP drosophila
Wild Type –
no nicotine
or nitric
oxide n = 20
APP
Drosophila–
no nicotine
or nitric
oxide n= 20
APP
Drosophila
–nicotine
n=20
APP
Drosophila
–nitric
oxide n=
20
Wild Type
–nicotine
n= 20
APP
Drosophila –
nitric oxide
and nicotine
n= 20
5 trials will be run, each for approximately 30 days per trial.
After all slides have been made they will be observed using the
fluorescent microscope and the images will be taken using a digital
camera and then using Photoshop the amyloid beta plaques will be
measured
Using SPSS statistical analysis will be done using an ANOVA
followed by a Scheffe Posthoc test with p< .05
Budget
Item Name
Vendor
Item #
#
Description
Price
Total Price
Drosophila Vials
Carolina Biology Catalog
173120
1
12 vials per order
$7.50
$7.50
Drosophila Food
Carolina Biology Catalog
173200
1
Formula 4-24® Instant
$6.95
$6.95
18.95
$18.95
Medium, Plain, 1 L
D. melanogaster, Large
Carolina Biology Catalog
172781
1
Large Culture: 70 to 100 flies
per order
PBS
Fisher Sciences Education
BP2438-4
5
A standard phosphate buffer
$95.56
$477.80
acetone
Carolina Biology Catalog
841481
5
Clear, colorless, flammable
$6.50
$32.50
liquid
methanol
Carolina Biology Catalog
874950
5
flammable
$5.75
$28.75
formalin
Carolina Biology Catalog
863463
5
flammable
$21.95
$109.75
xylene
Carolina Biology Catalog
898741
5
flammable
$10.60
$53.00
ethanol
Carolina Biology Catalog
861261
5
denatured with isopropanol
$7.95
$39.75
and methanol
distilled water
Carolina Biology Catalog
858621
5
Distilled Water
$7.25
$36.25
Serum Antibody Set
Carolina Biology Catalog
202102
1
Goat antisera containing
$99.00
$99.00
$11.75
$11.75
$8.50
$40
immunoglobulin
Disposable Lab Pack:
Carolina Biology Catalog
632920
1
50 Slides,50 Plastic
Coverslips,Lens Paper Booklet
i-Brite Plus
Neuromics
SF40000
1
anti-fade mounting media 10
ml
APP Drosophila
Not yet found
Phosphate Buffer Solution
Carolina Biology Catalog
2
746864
5
0.1 M Phosphate Buffer
Solution
Nitric Oxide
All Safe Industries
600-0054-000
1
25ppm Calibration Gas 58L.
$261.80
$261.80
Nicotine
Carolina Biology Catalog
877270
5
equal the amount of nicotine
$19.95
$99.70
found in one cigarette
Do Ability
• All materials are orderable from school
approved companies
• Drosophila can be kept alive with relative ease in
school lab
• Our school lab already has a microscope
necessary for observation
Bibliography
• Crag A. Michelli, William P Esler and W. Taylor Kimberley, Y-Secretase/presenilin inhibitors for
Alzheimer’s disease phenocopy Notch mutations in Drosophila, FASEB, 3/09/2003, volume 17,
pgs 79-81
• Crowther DC, Kinghorn KJ, Miranda E, Page R, Curry JA, et al. (2005) Intraneuronal Ab, nonamyloid aggregates and neurodegeneration in a Drosophila model of Alzheimer’s disease.
Neuroscience 132: 123–135.
• Harry EJ, Pogliano K, Losick R (1995) Use of immunofluorescence to visualize cellspecific gene
expression during sporulation in Bacillus subtilis. J Bacteriol
• 177, volume 12, pgs 3386-93.
• Gritta Tettweiler, Mathieu Miron, Mark Jenkins, Nahum Sonenberg, and Paul F. Lasko, Starvation
and oxidative stress resistance in Drosophila are mediated through the eIF4E-binding protein,
d4E-BP, Genes Dev, August 15 2005, volume 19, pgs 1840–1843.
• Isabell Greeve, Doris Kretzschmar, Jakob-Andreas Tscha¨pe, Anika Beyn, Claire Brellinger,
Michaela Schweizer, Roger M. Nitsch, and Rita Reifegerste1, Age-Dependent neurodegeneration
and Alzheimer-Amyloid Plaque Formation in Transgenic Drosophila, The Journal of
Neuroscience, April 21, 2004, volume 16, pgs 3899 –3906
• Koichi Iijima, Hsin-Ping Liu, Ann-Shyn Chiang, Stephen A. Hearn, Mary Konsolaki, Yi
ZhongDissecting the pathological effects of human Aβ40 and Aβ42 in Drosophila: A potential
model for Alzheimer's disease, PNAS, October 31, 2003, volume 101, pgs 6623–6628
• MUHAMMAD ARSHAD, MARIE PAULSSON, and PETR DEJMEK, Rheology of Buildup, of Acid
Casein Gels Breakdown, and Rebodying, March 3 1993, J Dairy Sci volume 76, pgs 331-3316
• P. Mpountoukas, A. Vantarakis, E. Sivridis and T. Lialiaria, Cytogenetic study in cultured human
lymphocytes treated with three commonly used preservatives, January 30 2008, Food and
Chemical Toxicology, volume 46, pgs 2390-2393
• Pogliano K, Harry E, Losick R (1995) Visualization of the subcellular location of
• sporulation proteins in Bacillus subtilis using immunofluorescence microscopy. Mol
• Microbiol, volume 18, pgs 459-70
• Yihong Ye and Mark E. Fortini Apoptotic Activities of Wild-type and Alzheimer’s Disease-related
Mutant Presenilins in Drosophila melanogaster, The Journal of Cell Biology, September 20 1999,
Volume 146, pgs 1351–1364