Shared Song Detector Neurons in Drosophila Male and Female Brains Drive Sex-Specific Behaviors: Shared Song Detector Neurons in Drosophila Male and Female Brains Drive Sex-Specific Behaviors (Current biology : CB (2019) 29 19 (3200-3215.e5) PII: S0960-9822(23)01049-7)

(Current Biology 29, 3200–3215.e1–e5; October 7, 2019) We repeated the pC2l silencing experiments (and controls) shown in Figures 5F, 5M–5O, S5F–S5H, and S6C–S6E. The new experiments were conducted in a modified behavioral rig (36 mm diameter, tiled with 16 microphones), and the UAS-TNT flies (w[^∗]; P{y[+t^∗] w[+mC]=UAS(FRT.stop)Ctet\tetX}VIE-19A/CyO) were re-ordered from the BDSC because it was discovered that the copy of this stock in the lab had become contaminated. We assayed paired courtship (pairs of single males and females); experimental or control flies courted (or were courted by) wild-type (NM91) flies. The genotypes of the experimental and control animals were as follows (and we verified the presence of TNT with PCR): Experiment (n = 21 male and n = 29 female): UAS > STOP > TNT/LexAop-FLP; dsx-LexA/R42B01-Gal4 Control (n = 17 male and n = 31 female): +/LexAop-FLP; dsx-LexA/R42B01-Gal4 In contrast to the original rig, the modified behavioral rig used 850 nm illumination (22 uW/mm² irradiance measured inside the chamber) for infrared (IR) video recordings, and the microphones were hand-painted with IR-absorbing dye (Epolin Spectre 160) to limit reflection artifacts in recorded videos. Further, the modified chambers were placed together with a white LED backlight within black acrylic enclosures to ensure constant lighting conditions across recordings. Video was recorded at 60 fps using the Motif recording system and API (loopbio GmbH, Austria). We observed different results with pC2l silencing in both males and females compared with the original published results, and the new versions of the figures are plotted below. The new results are now in line with what we had expected based on the published optogenetic activation and auditory responses of pC2l in males and females. We now find that silencing pC2l neurons in males strongly reduces song production (new Figure 5F) and mating success (new Figure S5H) and alters song, and silencing pC2l neurons in females reduces female slowing to pulse song (in particular, control flies show tuning for pulse IPI, whereas pC2l silenced females do not; new Figures 5M–5O). These new data now strongly support the conclusion that pC2l neurons in males are required for normal singing and that pC2l neurons in females are required for normal responses to male courtship song. We acknowledge Frederic Roemschied, Kyle Thieringer, and Yiqin Gao for new data collection and analysis.[Formula