The fly liquid-food electroshock assay (FLEA) reveals opposite roles for neuropeptide F in avoidance of bitterness and shock.

Abstract

Proper regulation of feeding is important for an organisms well-being and survival. Food intake in Drosophila can be determined in a number of ways, including by measuring the time a flys proboscis interacts with a food source in the fly liquid-food interaction counter (FLIC). Here, we show that electrical current flowing through flies during this interaction is aversive and leads to a reduction in food intake. Based on the FLIC, we engineer a novel assay, the fly liquid-food electroshock assay (FLEA), which allows for current adjustments for each feeding well. Using the FLEA, we show that both external incentives as well as internal motivational state can serve as drivers for flies to overcome higher current (electric shock) to obtain superior food. Unlike similar assays in which bitterness is the aversive stimulus for the fly to overcome, we show that current perception is not discounted as flies become more food-deprived. The FLEA is therefore a novel assay to accurately measure incentive motivation in Drosophila. Using the FLEA, we also show that neuropeptide F is required for proper perception or processing of an electroshock, a novel function for this neuropeptide involved in processing of external and internal stimuli. Significance StatementMany neuropsychiatric disorders, such as depression or addiction, are associated with alterations in motivated behavior. Assays measuring incentive motivation determine how driven an organism is to attain a goal, like food, or how attractive an incentive is. These tests require the animal to put effort into obtaining the reward, which can include physical work or overcoming an aversive stimulus. Such assays for Drosophila feeding have relied on flies overcoming bitterness to obtain their food. However, the perception of bitterness is discounted as flies become food deprived, confounding the interpretation. Here, we developed a novel assay that does not suffer from the same shortcomings and thus allows for more accurate assessments of incentive motivation in this widely used model organism.