Niederecker, Kirkpatrick, Attarian Final
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Transcript of Niederecker, Kirkpatrick, Attarian Final
The Effect of Caffeine on the Resistance to Heat in Drosophila melanogaster
Richard Niederecker, Kara Kirkpatrick, Samyar Attarian
Department of Biology SciencesSaddleback College
28000 Marguerite Parkway, Mission Viejo, CA 92692
Introduction• Drosophila melanogaster– Fruit fly
• Research benefits:– Short Lifespan– Distinct Developmental stages– Similarities to humans• Genome• Heat shock response
Lanneau, D., M. Brunet, E. Frisan, E. Solary, M. Fontenay, and C. Garrido. 2008. "Heat shock proteins: essential proteins for apoptosis regulation". Journal of Cellular and Molecular Medicine. 12 (3): 743-761.
Heat Shock Proteins (HSPs)• Historically drosophila cells
have been used to study heat shock proteins
• Protective function– Renature proteins
• Dependent Chaperone proteins– ATP
Example of Heat Shock Protein
Tissieres, A., Mitchell, H. K., and Tracy, U. (1974) Heat Shock Proteins: from Bacteria to Man, Cold Spring Proteins: from Bacteria to Man, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY. J. Mol. Bio. pg. 84, 389Beissinger, M., & Buchner, J. (1998). How chaperones fold proteins. Biol Chem, 379(3):245-59. http://www.ncbi.nlm.nih.gov/pubmed/9563819.Image by Jawahar Swaminathan and MSD staff at the European Bioinformatics Institute - http://www.ebi.ac.uk/pdbe-srv/view/images/entry/3hsc600.png, displayed on http://www.ebi.ac.uk/pdbe-srv/view/entry/3hsc/summary
Heat Shock Factors (HSFs)• Active HSFs bind to DNA to
stimulate production of HSPs• Monomer to Trimer• Trimer- active form• Trimer binds to DNA
Liou, S. (2010). The Heat-Shock Response. Miscellaneous Drugs. Huntington's Outreach Project for Education, at Sanford. Web. <http://web.stanford.edu/group/hopes/cgi-bin/hopes_test/the-heat-shock-response/>.
InducersNormal Cell Conditions(low concentration HSPs)
• Embryonic development• Hormonal stimulation• Cell differentiation• Cell cycle
– Cancer research
External Stressors (higher concentration HSPs)
• Oxidants• Heavy metals• Infectious diseases• Heat • Caffeine
• The Effect of Heat and Caffeine on the Development of Fruit Flies (Drosophila melanogaster) by Shiffman and Soliman (2014)
Shiffman, B. & Soliman, K. (2014). The Effect of Heat and Caffeine on the Development of Fruit Flies (Drosophila melanogaster). Department of Biological Sciences Saddleback College.
HypothesisDrosophila melanogaster consumption of caffeine will induce the production of HSPs, extending the lifespan of the flies in heat compared to heat alone.
Methods and Materials• 3 colonies– 120 flies each– Control and test
groups• Control groups
standard medium• Test groups
caffeinated medium• Incubated at 32C
• Medium production
Methods and Materials
• Two analyses – Pupae (1st colony)– Larvae (2nd and 3rd colonies)
• Extended caffeine exposure
• Incubated for full lifespan• Medium rehydrated
frequently– Increased caffeine
concentration– Dehydration of flies
One-tailed unpaired t-testp = 0.0047
Caffeinated Non-caffeinated0
1
2
3
4
5
6
7
8
9Pupa to Adult Lifespan
Fly Groups
Life
span
(day
s)
Caffeinated Non-caffeinated0
1
2
3
4
5
6
Larva to Pupa Lifespan
Fly Groups
Life
Spa
n (d
ays)
One-tailed unpaired t-testp = 1.45 x 10-13
Discussion and Conclusion
• Hypothesis rejected for all colonies
• Caffeine and heat are independent stressors
• Caffeine toxicity and heat• Conditioning
Muller, P. & Vernikos-Danellis , J. (1969). Effect of Environmental Temperature on the Toxicity of Caffeine and Dextroamphetamine in Mice. The Journal of Pharmacology and Experimental Therapeutics. Vol. 171 no 1 pg. 153-158Krebs, R. A., & Loeschcke, V. (1994). Costs and Benefits of Activation of the Heat-Shock Response in Drosophila melanogaster. Functional Ecology, 8(6), 730–737. http://doi.org/10.2307/2390232
Acknowledgements
Steve TehTony Huntley