Effects of prenatal ethanol exposure on acoustic characteristics of play fighting-induced ultrasonic vocalizations in juvenile rats

Abstract

Juvenile rats display rough-and-tumble playing with conspecifics (play fighting behavior) and produce 22 and 50 kHz ultrasonic vocalizations (USVs). The 22 kHz USV is considered to reflect negative emotionality such as anxiety, fear, and distress, whereas the 50 kHz USV is considered to reflect positive emotionality such as joy, happiness, and satisfaction. USV is a sensitive tool for measuring emotionality in socially interactive situations. However, effects of prenatal ethanol-exposure on the acoustic characteristics of play fighting-induced USVs have remained unclear. In Experiment I, we recorded USVs produced by prenatally ethanol-exposed rats during play fighting on postnatal days (PNDs) 40-42 and examined the acoustic characteristics of negative and positive emotion-induced USVs. In Experiment II, we examined the anxiety levels through elevated plus maze testing on PNDs 37-39 and frequencies of playful attacks on PNDs 43-45 in ethanol-exposed rats. Ethanol was administered to pregnant rats in three gradually increased concentrations between gestational days (GDs) 8 and 20. From GDs 14 to 20, ethanol-containing tap water at concentrations of 30% and 15% (v/v) was administered to the high- and low-ethanol groups, respectively. Tap water without added ethanol was given to the control group. On PNDs 40-42, three rats from the same sex and same ethanol concentration group but from different litters were placed together into a playing cage for play fighting. The high-ethanol male triads displayed elevations of 20-35 kHz USVs reflecting negative emotionality and reductions of 45-70 kHz USVs reflecting positive emotionality compared with both the low-ethanol and control male triads. The high-ethanol male triads had prominent elevations of 20-35 kHz USVs with durations longer than 200 ms, whereas the control male triads did not produce such 20-35 kHz USVs at all. There was no difference in USV acoustic characteristics among the female triads. In addition, the high-ethanol male rats exhibited greater anxiety levels and less frequencies of play fighting than the control male rats. Altogether, we conclude that prenatal exposure to ethanol enhances negative emotionality such as anxiety and, accordingly, 20-35 kHz USVs reflecting negative emotionality are produced with a marked decrease in play fighting, suggesting difficulties in social interactions with conspecifics.

Keywords: anxiety; ethanol; juvenile rats; play fighting; ultrasonic vocalization.