Extended Data Fig. 6: Pan-neuronal calcium dynamics in the song motor circuit during the singing of pulse and sine song.

a, Changes in calcium signals around song-type transitions for an example fly. Calcium signals were recorded from a dorsal part of the VNC. Left, average frames during pulse to sine transitions (n = 71 transitions). Images were averaged across z-planes and time. Right, average frames during sine to pulse transitions (n = 29 transitions). Scale bars, 50 μm. b, Population averaged changes in ΔF/F during song-type transitions for the recordings from the dorsal part of the VNC (n = 9 flies). Scale bars, 50 μm. c,d, Same as a,b but for the recordings from the medial part (n = 3 flies). (c) n = 171 and 62 pulse-to-sine and sine-to-pulse transitions, respectively. e, Schematic of the VNC region imaged in experiments where GCaMP was expressed using a pan-neuronal driver and CsChrimson was expressed with either the dPR1 or TN1A-2 driver. f, Pan-neuronal calcium imaging of the wing-related neuropils while activating dPR1. Frames were averaged across trials and flies (n = 6 trials for each irradiance; n = 7 flies), followed by further averaging across z-planes separately for dorsal and medial volumes of the VNC. The dashed circle in the image for the irradiance of 156 μW/mm² from the dorsal volume highlights a region that showed strong activation during dPR1 but not TN1A-2 stimulation. g, Same as f but for TN1A-2 (n = 6 trials for each irradiance; n = 6 flies). h, dPR1 neurons in the dorsal and medial volumes visualized with the dPR1 split-Gal4 line. Similar expression patterns were observed in three flies for each genotype. i, Same as h but for TN1A-2 neurons visualized with the TN1A-2 split-Gal4 line.