Ultrasoundgate cm16 cmpa

Ultrasonic sounds (see Fig. 2a for the outline of the processing) were detected using a condenser microphone (UltraSoundGate CM16/CMPA, Avisoft Bioacoustics, Berlin, Germany) designed for recordings between 10 and 200 kHz. The microphone was placed at a height of 20 cm from the floor and connected to an ultrahigh-speed ADDA converter BSA768AD-KUKK1710 and its software SpectoLibellus2D (Katou Acoustics Consultant Office, Kanagawa, Japan) with a sampling rate of 384 kHz (to measure 20–192 kHz). Recorded sounds were saved on a computer as wav files using SpectoLibellus2D. These sound files were analyzed with the GUI-based software USVSEG (implemented as MATLAB scripts) developed by us^{34 (link)}. Continuous sound signals with frequencies ranging from 40 to 160 kHz and durations ranging from 3 to 300 ms were analyzed and detected as syllables (USV calls). Devocalizing encounters were not tried because previous studies suggested that most vocalizations are produced by the resident male of the home cage and that the vocal contribution of the female is limited in the male–female courtship behaviors^{23 (link),30 (link),31 (link)} and the contribution of an intruder male is also limited in the male–male USVs^{23 (link)}.

Ultrasound recording was performed using a condenser microphone (UltraSoundGate CM16/CMPA, Avisoft Bioacoustics, Berlin, Germany) as previously reported [5] (link). When recording songs from adult males, the microphone was placed beside a cage (12×20×11 cm) with 6-cm diameter holes and covered with a 0.5-cm wide mesh. Ultrasonic sounds were recorded for 3 min when a male mouse encountered a devocalized female. Females were devocalized by sectioning of the inferior laryngeal nerve according to previous report [24] (link). Playback sounds consisted of 20 s of ultrasounds representing the strain character from 2 individuals each of B6 (number of syllables, 133 and 162) and BALB (number of syllables, 108 and 166). Similarly, playback sounds of ICR (number of syllables, 106) and KJR (number of syllables, 133; kindly gifted by Dr. Koide of the National Institute of Genetics, Mishima, Japan) were prepared. Figure S5 shows the principal components in 6 playback songs used in each behavior test. These sounds were repeatedly generated during the tests. Before each playback experiment, we played the song and simultaneously observed the sound pressure levels in the experimental box by using the same microphone kept away from the speaker, and adjusted the pressure levels.

In a soundproof chamber (O’Hara & Co.), ultrasonic sounds were detected by a condenser ultrasound microphone (UltraSoundGate CM16/CMPA, Avisoft Bioacoustics, Berlin, Germany) connected to an A/D converter (UltraSoundGate 116Hb, Avisoft Bioacoustics). Acoustic signals were recorded with an Avisoft SASLab Pro Recorder (Avisoft Bioacoustics) and transmitted to a sound analysis system (Avisoft SASLab Pro, Avisoft Bioacoustics).
To evaluate communication-related behaviour during postnatal stages, we monitored maternal separation-induced USVs as described previously^{34 (link)} with some modifications. Each pup was isolated from their mother and placed in a small cage containing fresh bedding. The number of USVs for 5 min was counted manually.
To examine adult vocal communication during different-sex interactions, we monitored courtship USVs as described previously^{39 (link)}. To induce courtship USVs in male mouse, an oestrous female C57BL/6J mouse was introduced into the male home cage. The number of USVs in a 5-min period was counted manually.
To further examine adult vocal communication during same-sex social interactions, we monitored USVs in the resident-intruder test as described previously^{28 (link)–30 (link)} with some modifications. An intruder C57BL/6J mouse was introduced to the resident home cage. The number of USVs in a 5-min period was counted manually.

To analyze the acoustic characteristics and create the playback files, we recorded the echolocation calls of the individual bats between 1900 and 2100 h when the bats were alert in a sound recording room (5 m long × 2 m wide × 2.7 m high). A single bat was perched in an experimental cage (60 × 60 × 60 cm) surrounded by sound-attenuating foam during the acoustic recording. An ultrasonic microphone (UltraSoundGate CM16/CMPA, Avisoft Bioacoustics, Berlin, Germany) was connected to an ultrasound recording interface with a sampling rate of 250 kHz and a 16-bit resolution, and every 60 s, the wave file generated was stored in a battery-powered notebook computer. The echolocation calls of each individual were recorded at a distance of 1 m from in front of the focal bat head. To ensure that the subsequent analysis and editing requirements were met, at least five 60 s sound files were recorded for each individual.