Figure 6. Activating RAm-projecting PAG neurons elicits USVs in male and female mice.

(A) (Left) Schematic showing the viral strategy used to express ChR2 in PAG-RAm neurons. Middle and right panels are confocal images of PAG (middle) and RAm (right), showing ChR2 labeling (green) and Fos expression (red) following USV production. (B) Spectrograms comparing USVs elicited by optogenetic activation of PAG-RAm neurons (left) and female-directed USVs (right) from the same animals. Top row shows example spectrograms from a male mouse, and the spectrograms in the bottom row are from a female. (C) Distributions of 5 acoustic parameters are shown for optogenetically-elicited USVs (blue) and female-directed USVs (gray) for N = 4 male and N = 3 female PAG_RAm-ChR2 mice (N = 2 females excluded from analysis that produced few female-directed USVs). Asterisks indicate acoustic parameters whose median value changed significantly in the same direction for 7/7 mice (p < 0.05, Mann Whitney U tests.) See also Fig. S2 and S6.