Extended Data Fig. 6: The role of Dh31 signaling in feeding to courtship transition.
From: A nutrient-specific gut hormone arbitrates between courtship and feeding
(a–b) Optogenetic activation of gut Dh31+ enteroendocrine cells promotes courtship and suppresses protein feeding. After a brief optogenetic stimulation of gut Dh31+ cells, 24-h starved males directly engage in courtship, unlike control flies that typically consumed amino acids (200 mM) before proceeding to courtship. Male flies were exposed to 660-nm LED light for 5 min and then transferred to the behavioral chamber for video recording for 30 min. Flies: Gut Dh31-GAL4: Dh31-GAL4, Tsh-GAL80. UAS: UAS-CsChrimson. Gut Dh31 CsChrimson flies contain all three transgenes. (a) Raster plots showing second-by-second behaviors of 5 representative males of each genotype. Behaviors: feeding (blue), courtship (orange) and copulation (black). (b) Fraction of time spent by each male on feeding and courtship. One-way ANOVA followed by Tukey’s test. n = 7 flies for each condition and genotype. (c–f) Dh31 in the gut and Dh31R in brain Crz+ neurons are required for males to transition from feeding to courtship. The Raster Plots show the second-by-second behaviors of 5 representative males. Gut Dh31-GAL4: Dh31-GAL4, Tsh-GAL80. Starved, 24 h. (c) Gut Dh31 knockdown reduced courtship activity and increased feeding in males ad libitum fed with standard fly food containing protein and carbohydrates. The fraction of time individual males spent on feeding and courtship is shown. Gut Dh31 KD flies had UAS-Dh31-RNAi and Gut Dh31-GAL4. One-way ANOVA followed by Tukey’s test. n = 7 flies for each condition. (d) Gut Dh31 knockdown increased feeding duration and eliminated courtship when food contained amino acids, but had no effect when food contained only sucrose. (e) Chemogenetic activation of gut Dh31-GAL4 cells. The ingestion of capsaicin-containing food induced courtship but food containing only sucrose had no effect. Gut Dh31 VR1 flies contained UAS-VR1 and Gut Dh31-GAL4. (f) Dh31R knockdown in brain Crz+ neurons eliminated the effect of amino acids on courtship, but not feeding. GAL4: Crz-GAL4, tub>GAL80>stop. UAS: otd-nls::flp, UAS-Dh31R-RNAi. Brain Dh31R KD flies had all four transgenes.
