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Updated: May 3, 2026

Determining Ultrasonic Vocalization Preferences in Mice using a Two-choice Playback Test
Published on: September 3, 2015
Developmental social environment imprints female preference for male song in mice
Akari Asaba1, Shota Okabe1, Miho Nagasawa1
1Companion Animal Research, School of Veterinary Medicine, Azabu University, Sagamihara, Kanagawa, Japan.
Background:
Sexual imprinting is important for kin recognition and for promoting outbreeding, and has been a driving force for evolution; however, little is known about sexual imprinting by auditory cues in mammals. Male mice emit song-like ultrasonic vocalizations that possess strain-specific characteristics.
Objectives:
In this study, we asked whether female mice imprint and prefer specific characteristics in male songs.
Methods And Findings:
We used the two-choice test to determine the song preference of female C57BL/6 and BALB/c mice. By assessing the time engaged in searching behavior towards songs played back to females, we found that female mice displayed an innate preference for the songs of males from different strains. Moreover, this song preference was regulated by female reproductive status and by male sexual cues such as the pheromone ESP1. Finally, we revealed that this preference was reversed by cross-fostering and disappeared under fatherless conditions, indicating that the behavior was learned by exposure to the father's song.
Conclusions:
Our results suggest that female mice can discriminate among male song characteristics and prefer songs of mice from strains that are different from their parents, and that these preferences are based on their early social experiences. This is the first study in mammals to demonstrate that male songs contribute to kin recognition and mate choice by females, thus helping to avoid inbreeding and to facilitate offspring heterozygosity.
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