The effect of playback of 22-kHz and 50-kHz ultrasonic vocalizations on rat behaviors assessed with a modified open-field test.

Juvenile and adult rats emit two affectively different types of ultrasonic vocalizations (USVs), namely aversive 22-kHz and appetitive 50-kHz USVs. Aversive 22-kHz USVs are considered to be alarm calls that communicate negative affective states to conspecific receivers. Although the alarming effects of playback of 22-kHz USVs were reported recently, behavioral data showing those effects are still not abundant. Appetitive 50-kHz USVs are considered to communicate positive affective states to conspecific receivers, to pace and coordinate social behavior. In line with this, playback of 50-kHz USVs has been found to initiate behavioral activation and induce approach behavior in receiver rats. However, most of these playback studies have used male 50-kHz USVs; thus, it seems to remain unclear whether female 50-kHz USVs exert a similar social attractant effect on male rats. To investigate these issues, we performed modified open-field tests, during which USVs were continuously presented for 15 min to male receivers. In these tests, if negative affective changes are evoked in subject rats, the time spent in the open arena decreases, while the time spent on defensive behaviors increases. In contrast, when positive affective changes are evoked, the opposite phenomenon is observed. Playback of male aversive 22-kHz USVs induced anxiety-related defensive responses in receivers. However, playback of female appetitive frequency-modulated (FM) 50-kHz USVs increased opposite, appetitive pattern of exploratory behavior with increased exploration. The results indicate that playback of male aversive 22-kHz and female appetitive 50-kHz USVs might induce behavioral responses probably associated with negative and positive affective states in male rats, respectively, suggesting the validity of rat USVs as an animal model of vocal communication of emotion.