Extended Data Fig. 10: Circuit and intrinsic properties of PAGVGAT neurons support USV persistence.
From: Flexible scaling and persistence of social vocal communication
a, b, USVs emitted during photostimulation of LPOAVGAT/ChR2 cells with injection of saline as control (grey) or CNO (purple) to inhibit PAGVGAT/hM4Di neurons on alternative days. a, Number of USVs emitted with 1 Hz 5s light stimulation. b, Raster plot of USVs emitted during and after photostimulation (blue bars). Each row is 30 s of a 230-s trial aligned to light stimuli applied every 40 s. Data are mean ± s.e.m. n = 3, 6 trials per mouse per test day. ****P < 0.0001, paired Wilcoxon test, two sided. ns, not significant (P > 0.05). c, Representative image of co-expression of hM4Di-tdTomato with VGAT-ZsGreen cells in the PAG. Scale bar, 200 μm. n = 2 mice, 2 sections per mouse collected. d, Number of USVs emitted during increased frequency photostimulation in LPOA of male mice expressing LPOAVGAT/ChR2 and PAGVGAT/hM4Di and injected with CNO or saline on alternative days. n = 3, 36 trials per condition. Data are mean ± s.e.m. ****P < 0.0001, Wilcoxon test, two sided. e, Experimental design to express control virus (tdTomato) in the PAG of VGAT-Cre mice (Fig. 4e, f). f, Number of USVs emitted by photostimulating LPOAVGAT cells under either CNO or saline conditions in mice expressing FLEX-tdTomato viral control (PAGVGAT/tdTomato) (control). n = 3, 18 trials per condition. Data are mean ± s.e.m. ns, not significant (P > 0.05), Wilcoxon test, two sided. g, Raster plot of USV bouts emitted during either CNO or saline test days under different stimulation frequency. Note that Drosophila courtship songs show similar feature separation across the circuit with songs evoked by pIP10 neurons tightly locked to stimulation (like the PAG USV-gate neurons) compared to calling generated by P1 neurons that persists beyond stimulation (as with the LPOAESR1 neurons)53. This suggests diverse social species use general circuit strategies to maintain persistent auditory responses that outlast the detection of sensory information.
