Ultrasoundgate 416 hr 1.1 hardware

Ultrasonic vocalizations (USVs) were recorded from both male and female pups isolated from their mothers at P4, during the daylight period of the light/dark cycle. Dams and their litters were acclimated for 30 min in the test room. Each pup was removed from the cage containing its mother and littermates and placed in a clean plastic container in a wooden sound-attenuating recording chamber. Each pup was first acclimated in the recording chamber for 30 s then recorded for 10 min. Recordings were acquired using an UltraSoundGate CM16/CMPA condenser microphone (Avisoft Bioacoustics) positioned at a fixed height of 8 cm above the pups, and were amplified and digitized (sampled at 16 bits, 250 kHz) using UltraSoundGate 416 hr 1.1 hardware and Avisoft-RECORDER software (Avisoft Bioacoustics). The data were transferred to Avisoft-SASLab Pro (version 5.2) to analyze spectrograms of vocalizations with settings of 0% overlapping FlatTop windows, 100% frame size, and 256 points fast Fourier transform (FFT) length. The following measures were recorded for each group: number of USV calls, mean duration of USV calls, and mean peak frequency. Recordings were performed with the experimenter blinded to mouse genotypes.

USVs were recorded at P4 for both male and female pups isolated from their mothers during the daylight period of the light/dark cycle. Dams and their litters were acclimated to the test room for 30 min. Each pup was removed from the cage containing its mother and littermates and placed in a clean plastic container in a wooden sound-attenuating recording chamber. Each pup was first acclimated to the recording chamber for 30 s (sec) then recorded for 10 min. Recordings were acquired using an UltraSoundGate CM16/CMPA condenser microphone (Avisoft Bioacoustics) positioned at a fixed height of 8 cm above the pups, and were amplified and digitized (sampled at 16 bits, 250 kHz) using UltraSoundGate 416 hr 1.1 hardware and Avisoft-RECORDER software (Avisoft Bioacoustics). The data were transferred to Avisoft-SASLab Pro (version 5.2) to analyze spectrograms of vocalizations with settings of 0% overlapping FlatTop windows, 100% frame size, and 256 points fast Fourier transform (FFT) length. The following measures were recorded and analyzed for each group: number of USV calls, mean duration of USV calls, mean amplitude, mean peak frequency, maximum peak frequency, mean interval between USV calls, and the latency to call.