Abstract
Animals, from insects to humans, exhibit obvious diurnal rhythmicity of feeding behavior. Serving as a genetic animal model, Drosophila has been reported to display feeding rhythms; however, related investigations are limited due to the lack of suitable and practical methods. Here, we present a video recording-based analytical method, namely, Drosophila Feeding Rhythm Analysis Method (dFRAME). Using our newly developed computer program, FlyFeeding, we extracted the movement track of individual flies and characterized their food-approaching behavior. To distinguish feeding and no-feeding events, we utilized high-magnification video recording to optimize our method by setting cut-off thresholds to eliminate the interference of no-feeding events. Furthermore, we verified that this method is applicable to both female and male flies and for all periods of the day. Using this method, we analyzed long-term feeding status of wild-type and period mutant flies. The results recaptured previously reported feeding rhythms and revealed detailed profiles of feeding patterns in these flies under either light/dark cycles or constant dark environments. Together, our dFRAME method enables a long-term, stable, reliable, and subtle analysis of feeding behavior in Drosophila. High-throughput studies in this powerful genetic animal model will gain great insights into the molecular and neural mechanisms of feeding rhythms.
Highlights
Feeding rhythm has recently been found to play a critical role in animal health
We developed a new analytical method, dFRAME, which is simple, stable, robust, and reliable for investigating the feeding rhythms of fruit flies
For dFRAME: Feeding Rhythm in Drosophila studying feeding rhythms in a long term, dFRAME is ideal to perform high-throughput screen and explore new behavioral patterns, and ARC can be used to determine the amount of food intake
Summary
Feeding rhythm has recently been found to play a critical role in animal health. It is vital for animals to synchronize their feeding behavior to both internal biological clock and external environmental conditions, while the underlying mechanisms remain to be elucidated. With powerful tools for genetic and neural manipulation, Drosophila melanogaster serves as an excellent model for investigating the molecular and neural mechanisms underlying circadian rhythms (Franco et al, 2018). Earlier studies in Drosophila uncovered the molecular mechanisms controlling internal clock, which are evolutionarily conserved from insects to mammals (Yu and Hardin, 2006). The sleep rhythm and the role of light entrainment have been well studied in flies (Shafer and Keene, 2021). Studies on feeding rhythms remain limited, largely because of the limitation of available analytical methods of fly feeding
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