Behavioural response of female Lewis rats toward 31-kHz ultrasonic calls

Rodent ultrasonic vocalisations can be used to assess social behaviour and have attracted increasing attention. Rats emit 50-kHz and 22-kHz calls during appetitive and aversive states, respectively. These calls induce behavioural and neural responses in the receiver by transmitting the internal states of the rats, thus serving communicative functions. Recently, we discovered that female Lewis rats emit 31-kHz calls under social isolation and inequality conditions; however, the biological significance of 31-kHz calls remains unknown. In the present study, we conducted three playback experiments to examine the behavioural effects of 31-kHz calls. In the first experiment, Lewis female rats were exposed to four types of sound: 22-kHz, 50-kHz, 31-kHz calls, and environmental noise. As a result, rats stayed significantly longer in the area with a sound-producing speaker, regardless of the sound type, than in the silent speaker area. The duration spent around the sound-producing speaker was particularly extended during the 50-kHz or 31-kHz call playback, compared to the environmental noise or 22-kHz call playback. In the second experiment, rats were exposed to refined versions of sound stimuli that were synthesised to preserve prominent frequency components while removing background noise from original calls. Rats significantly preferred to stay around the speaker for the synthesised 50-kHz and 31-kHz sounds, but not for the synthesised 22-kHz sound. However, in the third experiment, additional 31-kHz sound synthesised from calls emitted by a different rat did not elicit a significant preference for the source side. These results suggest that the rats paid attention to the 31-kHz call, although it is plausible that acoustic variability in the 31-kHz USV may affect their approach behaviour.