The Effects Of Nitric Oxide On The Development Of Alzheimer's DiseaseCaroline Hsiao

Need

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Need

http://msnbcmedia1.msn.com/i/msnbc/Components/Art/HEALTH/070319/AP_ALZHEIMERS.gif

Plaques

PlaquesTangles in Neuron

Alzheimer’s Disease

http://pampee.20gig.com/wp-content/uploads/2009/04/alzheimers-disease.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

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http://www.inchem.org/documents/jecfa/jecmono/40abcj11.gif

Oxidative Stress•Cigarette smoke is used as an example

shown here

http://healthy-lifestyle.most-effective-solution.com/wp-content/uploads/2007/10/oxidative-stress.jpg

Age-Dependent Neurodegeneration and Alzheimer-Amyloid Plaque Formation in Transgenic Drosophila

Greeve, and Kretzschmar,

Dissecting the pathological effects of human A40 and A42 in Drosophila: A potential model for Alzheimer’s disease

By: Iijima and Liu

Investigation of Free Radicals in Cigarette Mainstream Smoke M. Ghosh and P. Ionita

Purpose

Alternate: Nitric Oxide will cause an increase in the development of

Alzheimer's symptoms in drosophila for all groups tested.

Null Hypothesis: there will be no change in any of these groups at all.

The purpose of this experiment was to find if nitric oxide have any effect on

the development of Alzheimer's disease.

Experimental DesignProtocols 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 nitric oxide n = 20

APP Drosophila–no nitric oxide n= 20

APP Drosophila –nitric oxide for 1 minute n= 20 (6 days old)

Wild Type-nitric oxide n= 20 (6 days old)

5 trials will be run, each for approximately 12-13 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 total amount of 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

Wild Type–nitric oxide for 1 minute n= 20

APP Drosophila –nitric oxide for 5 minute n= 20 (6 days old)

Wild Type–nitric oxide for 1 minute n= 20

BudgetItem 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

Medium, Plain, 1 L

$6.95 $6.95

D. melanogaster, Large Carolina Biology Catalog 172781 1 Large Culture: 70 to 100 flies

per order

18.95 $18.95

PBS Fisher Sciences Education BP2438-4 5 A standard phosphate buffer $95.56 $477.80

acetone Carolina Biology Catalog 841481 5 Clear, colorless, flammable

liquid

$6.50 $32.50

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

and methanol

$7.95 $39.75

distilled water Carolina Biology Catalog 858621 5 Distilled Water $7.25 $36.25

Serum Antibody Set Carolina Biology Catalog 202102 1 Goat antisera containing

immunoglobulin

$99.00 $99.00

Disposable Lab Pack: Carolina Biology Catalog 632920 1 50 Slides,50 Plastic

Coverslips,Lens Paper Booklet

$11.75 $11.75

i-Brite Plus Neuromics SF40000 1 anti-fade mounting media 10

ml

APP Drosophila Not yet found 2

Phosphate Buffer Solution Carolina Biology Catalog 746864 5 0.1 M Phosphate Buffer

Solution

$8.50 $40

Nitric Oxide All Safe Industries 600-0054-000 1 25ppm Calibration Gas 58L. $261.80 $261.80

Do Ability•All materials are orderable from school

approved companies

•Drosophila can be kept alive with relative ease in school lab

•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, non-amyloid 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