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Rapid and Chronic Tolerance of Ethanol in Drosophila Melangastor By: Paul Mascali, Anthony Anastasio Abstract Alcohol is the most abused drug in the world. 30 percent of Americans have reported having some form of alcohol use dependence. Drosophila is one of the only model organisms that can metabolize ethanol or the pure form of alcohol. While the ethanol is being metabolized drosophila build up a tolerance. There are to forms of tolerance, rapid and chronic tolerance. These different forms are established by the exposures of the ethanol. Both can be measured in comparison to show flies that have built up a tolerance can take in more ethanol than the flies that have never been exposed to ethanol. Need Graph A Alcohol has the largest death rate out of all other drugs Graph B disorders caused by alcohol have increased over the past 6 years Knowledge Base http://chidiet.com/images3/banana1.jpg http://www.imsb.ethz.ch/researchgroup/hafene/researchprojects/Drosophila.jpg Drosophila relate to humans on a genetic level Ethanol can be found in the fermentation of fruits Knowledge Base Scale defined by Sullivan 2000 0 normal behavior 1 continuous grooming 2 Loss of flight 3 slow locomotion 4 backward locomotion 5 bouncing off the walls 6 sever body tremors 7 death Literature Review Rapid and chronic: two distinct forms of ethanol tolerance in Drosophila Urizar, 07 Rapid tolerance-defined as the attenuated response to ethanol that occurs from a single exposure and after elevated blood levels from the initial exposure Chronic tolerance-when they are exposed for long amount of time to small concentration of the drug. Literature Review Rapid and Chronis: Two Distinct Forms of Ethanol Tolerance in Drosophila Ulrike Heberlein Rapid Tolerance 75% alcohol for 15 minutes 100% alcohol for 7 minutes Chronic tolerance 10% alcohol for 4 hrs-2 days 10% alcohol for 24 hours Literature Review Drosophila Homer Is Required in a Small Set of Neurons Including the Ellipsoid Body for Normal Ethanol Sensitivity and Tolerance Nancy L. Urizar (alcohol) (alcohol) (alcohol) Literature Review Phillips, 06 ETHANOL TOLERANCE IN TEMPERATE DROSOPHILA Lower the Temperature the flies had increased levels of lipid biosynthetic enzymes such as phospholipase D which betters tolerance Hypothesis • null hypothesis-the drosophila that are never exposed to ethanol will have the weaker tolerance • alternative hypothesis-based on the findings of Ulrike Heberlein flies with chronic tolerance will have a stronger tolerance Purpose The purpose of the study is to show that tolerance has an affect on the metabolism of alcohol in drosophila melagastor. Methodology/ do-ability Air pump Ethanol beaker Testing chamber http://images.google.com/imgres?imgurl=http://bp3.blogger.com/_q3QBSc SSHhw/Rho69SisEI/AAAAAAAAF7s/dTLsMme2ZHQ/s320/PMA%2BAldrich.jpg&img refurl=http://curlyarrow.blogspot.com/2008/03/lets-talk-about-tlcs-part3.html&h=320&w=144&sz=13&hl=en&start=2&um=1&tbnid=THmY2aSgmI LpgM:&tbnh=118&tbnw=53&prev=/images%3Fq%3D100%2525%2Bethan ol%2Bcontainer%26um%3D1%26hl%3Den%26rls%3DSUNA,SUNA:200602,SUNA:en%26sa%3DN (Taken by author) Methodology/ do-ability Pipe controlling flow of ethanol gas Inebriometer Ethanol Beaker Air Pump Funnel Layers http://blog.wired.com/wiredscience/images/2007/11/01/inebriometer.gif Test to determine when ethanol intoxication takes place (Each group was fresh flies never exposed to ethanol) Wild Type Drosophila Repeat after each death of test subjects for 6-7 different trials Flies that built up a rapid tolerance Flies never exposed to alcohol Expose flies to 20 proof ethanol for 24 hours Expose flies to 200 proof ethanol for 7 minutes once a week Repeat each week Put flies in inebriometer several hours after exposure Flies that built up a chronic tolerance Count the number of levels the flies fall through the Inebriometer Put flies in inebriometer 24 hours after exposure After all data is collected it is placed into SPSS for data analyzes Bibliography • • • • • • • • • • • • • Tabakoff,B; Conrnell, N; &Hoffman, P.L. Alcohol tolerance. Annals of Emergency Medicine 15(9):1005-1012, 1986 Kathryn Phillips, Ethanol Tolerance in Temperate Drosophila, Journal of Experimental Biology 209, September 19, 2006 Heberlein Ulrike. Molecular Genetic Analysis of Ethanol Inoxication in Drosophila Melanogaster. Aug. 2004 Natural selection protected some east Asian Population form Alcoholism Scinece Daily 2003 Jama and Archives journals (2007) Almost one-third fo Adults Report Having some Form of Alcohol use problem during their lifetime. Science Daily Sullivan Ashburner Hawley, Drosohila Protocols, Cold Spring Habor Laboratory Press, 1996 Mumenthaler, Martin Gender Differences in Moderate drinking effects Alcohol Health & Research World Chien Samson Homophila:human disease gene cognates in Drosophila Nucleiec Acids Research Steven Gans Fruitflies Used to Examin Alcohol Tolerance 2000 Mumenthaler Martin Gender Differences in Moderate Drinking Effects Alcohol Health &Research World Winberg Jan Olof Drosophila Melanogsater Alcohol Dehydrogenase: Product-Inhibition 1994 Ulrike Heverlein Molecular GeneticAnalysis of Ethanol Intoxication In Drosophila Melangaster 2004 Department of Anatomy and Program in Neuroscience Morozava Transcriptional response to alcohol exposure in Drosophila Melanogaster; Genome Biology 2006