Rapid and Chronic Tolerance of Ethanol in Drosophila

Melangastor

By: Paul Mascali, Anthony Anastasio

AbstractAlcohol 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 AGraph B

Alcohol has the largest death rate out of all other drugs

disorders caused by alcohol have increased over the past 6 years

Knowledge Base

http://www.imsb.ethz.ch/researchgroup/hafene/researchprojects/Drosophila.jpg

http://chidiet.com/images3/banana1.jpg

Drosophila relate to humans on a genetic level

Ethanol can be found in the fermentation of fruits

Knowledge Base

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

Scale defined by Sullivan 2000

Literature Review

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.

Urizar, 07

Rapid and chronic: two distinct forms of ethanol tolerance in Drosophila

Literature ReviewRapid and Chronis: Two Distinct Forms of Ethanol Tolerance in Drosophila

Ulrike Heberlein

Rapid Tolerance

Chronic tolerance

75% alcohol for 15 minutes

100% alcohol for 7 minutes

10% alcohol for 4 hrs-2 days

10% alcohol for 24 hours

Literature Review

Drosophila Homer Is Required in a Small Set of NeuronsIncluding the Ellipsoid Body for Normal Ethanol Sensitivityand 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

(Taken by author)

Ethanol beaker

Testing chamber

Air pump

http://images.google.com/imgres?imgurl=http://bp3.blogger.com/_q3QBScSSHhw/R-ho69SisEI/AAAAAAAAF7s/dTLsMme2ZHQ/s320/PMA%2BAldrich.jpg&imgrefurl=http://curlyarrow.blogspot.com/2008/03/lets-talk-about-tlcs-part-3.html&h=320&w=144&sz=13&hl=en&start=2&um=1&tbnid=THmY2aSgmILpgM:&tbnh=118&tbnw=53&prev=/images%3Fq%3D100%2525%2Bethanol%2Bcontainer%26um%3D1%26hl%3Den%26rls%3DSUNA,SUNA:2006-02,SUNA:en%26sa%3DN

Methodology/ do-ability

Ethanol Beaker

Air Pump

Inebriometer

Funnel Layers

Pipe controlling flow of ethanol gas

http://blog.wired.com/wiredscience/images/2007/11/01/inebriometer.gif

Expose flies to 200 proof ethanol for 7 minutes once a week

Expose flies to 20 proof ethanol for 24 hours

Count the number of levels the flies fall through the Inebriometer

After all data is collected it is placed into SPSS for data analyzes

Repeat each week

Repeat after each death of test subjects for 6-7 different trials

Test to determine when ethanol intoxication takes place (Each group was fresh flies never exposed to ethanol)

Wild Type Drosophila

Flies that built up a rapid tolerance

Flies that built up a chronic tolerance

Flies never exposed to alcohol

Put flies in inebriometer several hours after exposure

Put flies in inebriometer 24 hours after exposure

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