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Transgenerational Effects of Parental Ethanol Consumption on Offspring Anxiety Behavior in Response to Stress Open Access

In the wild, Drosophila melanogaster is exposed to ethanol as a result of yeast fermentation of the sugar that is found in its natural food source, fruit. Previous work in our lab found that diet-induced ethanol exposure created heritable epigenetic changes in subsequent generations. Here, I investigated whether this epigenetic change was correlated with anxiety. To test this I fed a parental generation of Drosophila melanogaster alcohol using Capillary Feeders (CAFEs) of liquid media containing sugar, water, and ethanol (A treatment). A control group was also fed using CAFEs, with liquid media that did not contain alcohol (N treatment). After three days of this treatment, I bred the alcohol-fed males and females (AxA) and the control males and females (NxN) to produce the F1 generation. I exposed the F1 generation to variable chronic stressors over six days, consisting of alternating heat shock at 34 degrees for 3 hours, cold shock at 4 degrees for 3 hours, and starvation (48 hours without food). I then measured the anxiety levels of these offspring using two different assays: a light-dark box (LDB) and a forced swim test (FST). These results were compared to the positive control, represented by a strain of Drosophila melanogaster that was given food (consisting of water, sugar, agar, and yeast) without the use of CAFEs. All behavioral trials were run with care given to isolating the stress response (indicative of anxiety behavior) from any other behavioral interactions. This included running male and female trials separately, aging flies for the same number of days before the trial runs, running trials at the same time of day, and taking precautions to minimize disruptions so to maintain a controlled testing environment throughout all assays.

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