Neuroscience JoVE (Open Access) Citable · DOI

Eliciting and Analyzing Male Mouse Ultrasonic Vocalization (USV) Songs

Name: Eliciting and Analyzing Male Mouse Ultrasonic Vocalization (USV) Songs
Uploaded: 2026-06-04
Duration: 8 min 44 s
Description: Mice produce ultrasonic vocalizations (USVs) in a variety of social contexts throughout development and adulthood. These USVs are used for mother-pup ...

DOI: 10.3791/54137-v

What you'll learn

✓ Consistently elicit and record male mouse ultrasonic vocalizations (USVs) under controlled conditions
✓ Extract and quantify acoustic features and syllabic syntax from USV recordings
✓ Analyze female preference for male song variants using playback experiments
✓ Apply USV analysis as a behavioral readout for neuropsychiatric phenotyping

Protocol

Mice produce a complex multisyllabic repertoire of ultrasonic vocalizations (USVs). These USVs are widely used as readouts for neuropsychiatric disorders. This protocol describes some of the practices we learned and developed to consistently induce, collect, and analyze the acoustic features and syntax of mouse songs.

Difficulty

intermediate

Total time

~1–2 hours per recording session; full experiment with playback analysis ~3–4 days

Model organism

Mouse (male and female, species not specified)

Steps

Prepare test mice for recording

Acclimate male mice to the testing room and handling procedures to habituate them prior to USV elicitation.

▶ 00:49

Prepare female mouse as stimulus

Set up female stimulus mice (anesthetized or restrained) to elicit male courtship ultrasonic vocalizations during the recording session.

▶ 01:50

Record ultrasonic vocalizations

Position ultrasonic microphones and recording equipment to capture male USVs during interaction with the female stimulus; document raw acoustic data.

▶ 03:23

Conduct playback experiment

Replay recorded male USV variants to naive females and measure behavioral/preference responses to assess song attractiveness.

▶ 04:38

Analyze song diversity and female preference

Quantify acoustic parameters (frequency, duration, syllable structure) and correlate with female behavioral preference data to characterize song phenotype.

▶ 06:48

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