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Drunk Drosophila: Analysis of Long and Short-Term Exposure of Apterous (wingless) Drosophila Melanogaster to Ethanol

Vanessa Akwada and Aina Muhd Shahrulmiza� BioBus. Inc., Harlem, New York

Alcohol and substance abuse are afflictions that have detrimental effects on its abusers. Environmental factors such as availability and societal influences are believed to affect the individual response to alcohol abuse. Drosophila melanogaster -the fruit fly- has been previously used as a model for addiction. Both humans and fruit flies demonstrate similar behaviors when intoxicated with different percentages of ethanol. In its natural environment, Drosophila melanogaster encounters high levels of ethanol. Some fruit flies subsist on rotten fruit that can contain over 5% ethanol. It has been previously shown that wild type Drosophila prefer ethanol-rich foods over ethanol after short-term exposure to ethanol. We sought to use the mutant Apterous (wingless) Drosophila and a new system of analysis to recapitulate and expand on these data. Experiments were conducted to determine Drosophila’s preference to different methods of ethanol presentation, such as ethanol-infused foods or substrates when they were starved and not starved. The results from these series of experiments led us to conclude that Apterous (wingless) Drosophila prefer ethanol-rich foods after short-term and long-term exposure regardless of concentration of ethanol.

Hypothesis: The Apterous (ap) mutant of Drosophila melanogaster will exhibit behavioral differences when exposed to ethanol or ethanol-rich foods. The duration of exposure would affect preference.

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1. Starved and non-starved wingless fly populations were used in each experiment. The starved flies were deprived of food for 6 hours prior to running the experiment.
2. Choice Chambers were used to test various conditions for Drosophila’s preference for ethanol-rich food with different percentages of ethanol:�*Our data was derived from several experimental conditions:
1. Establishing Experimental Parameters:
1. Colored lights vs. white light (Red vs. white light, blue vs. white light, green vs. white light, no light vs. white light)
2. Short Term (15 mins): Banana Infused EtOH vs. Red Light + 5% EtOH
3. Short Term (15 mins): Banana Infused EtOH v. Red Light (no alcohol)
2. Long Term 5% EtOH exposure (expose flies to alcohol in their environments for various duration of time + test their preferences):
• 5 Days
• 18 Days
• 26 Days
• 33 Days
• 39 Days
• 60 Days
3. Long Term 20% EtOH exposure:
• 1 Day
• 3 Days
• 4 Days
• 5 Days
3. Preference for the control vs. the experimental conditions were determined in 1 minutes intervals for 10 minutes, and then again at 15 minutes. Trials were done in triplicate.

Results

Conclusion and Future Directions

Conclusion:

• There is no difference between short term and long term exposure with non-starved and starved flies
• Flies exposed to high concentrations of ethanol demonstrate a change in morphology
• Flies exposed to 5% ethanol have a shorter walk frequency but demonstrate a similar maximum walk velocity as the control
• Flies exposed to 20% ethanol have similar walk frequencies but drastic difference in maximum walk velocity compared to the control

Future Directions:

• Separate male and females and test their preference
• Utilize mutants flies that can’t smell banana and test their preference
• Build a better choice chamber and code a program that will allow a camera to take pictures every minute → will improve data collection

Introduction

Works Cited

1. Devineni AV, Heberlein U (2009) Preferential Ethanol Consumption in Drosophila Models Features of Addiction Curr Biol. Dec 29; 19(24): 2126–2132.
2. Vang LL, Medvedev AV, Adler J (2012) Simple Ways to Measure Behavioral Responses of Drosophila to Stimuli and Use of These Methods to Characterize a Novel Mutant. PLoS ONE 7(5): e37495. doi:10.1371/journal.pone.0037495
3. Kaun, K. R., Devineni, A. V., & Heberlein, U. (2012). Drosophila melanogaster as a model to study drug addiction. Human Genetics, 131(6), 959–975. http://doi.org/10.1007/s00439-012-1146-6
4. Guarnieri DJ, Heberlein U (2003) Drosophila melanogaster, a genetic model system for alcohol research, International Review of Neurobiology, 199-228.
5. Heberlein U, Wolf FW, Rothenfluh A, Guarnieri DJ (2004) Molecular Genetic Analysis of Ethanol Intoxication in Drosophila Melanogaster, Integrative and Comparative Biology, 44(4):269-74. doi: 10.1093/icb/44.4.269.

Methods

Materials

• CAROLINA™ Choice Chambers
• Petri Dishes
• Plastic/disposable mL-pipettes
• 20% ethanol and 5% ethanol
• Apterous (ap) mutant wingless flies (new flies used for every trial)
• Cotton balls
• Timer
• Funnel
• Fresh fly food (Yeast for adult flies, Drosophila Media for larvae)
• 1 fresh banana
• Red flashlight
• Cardboard box

Manhattan Center for Science and Mathematics

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Baruch College Campus High School

Richard Mann’s Lab Data

Key for Figures 1-3

Figure 1: Graphs display the average percentage of non-starved (A) and starved flies (B) in choice chambers that contain banana infused 5% ethanol versus red light and pure 5% ethanol.

Figure 4: Depiction of adult flies. Morphology of control fly versus long term fly exposure exposed to 20% ethanol. The fly on the left is the control fly, and the fly on the right is the experimental fly after long term exposure to 20% ethanol.

Figure 2: Graphs display the average percentage of starved flies in choice chambers that contain banana infused 5% ethanol versus red light and pure 5% ethanol. Flies were exposed to 5% ethanol for 5 days (A) and 18 days (B) prior to the choice chamber experiment.

Figure 3: Graphs display the average percentage of non-starved (A,C) and starved flies (B,D) in choice chambers that contain banana infused 5% ethanol versus red light and pure 5% ethanol. Flies were exposed to 5% ethanol for 26 and 60 days prior to the choice chamber experiment.

Figure 5: Graphs display the average percentage of non-starved (A) and starved flies (B) in choice chambers that contain banana infused 20% ethanol versus red light and pure 20% ethanol. Flies were exposed to 20% ethanol for 1 day prior to the choice chamber experiment.

Figure 6: The Mann lab has developed a novel technique to analyze fly mobility. Through the use of high-speed imaging of live flies, we can quantify walk frequency, average walk velocity, and average total distance during a 3 minute exposure time.