Video 2. Optogenetic activation of Amg_C/M-PAG neurons causes no obvious behavioral effects in the absence of a social partner.

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An example male mouse with ChR2 expression in Amg_C/M-PAG neurons is shown alone in a chamber with no social partner. Optogenetic activation of Amg_C/M-PAG neurons with pulses of blue light does not elicit ultrasonic vocalization (USV) production or any other obvious behavioral response. Video is shown at the top, a spectrogram (bottom) showing the audio recording is synchronized to the video, and pitch-shifted audio (80 kHz to 5 kHz transformation) is included to place any USVs that may have occurred within the human hearing range.