Eliciting and Analyzing Male Mouse Ultrasonic Vocalization (USV) Songs

Abstract

Mice produce ultrasonic vocalizations (USVs) in a variety of social contexts throughout development and adulthood. These USVs are used for mother-pup retrieval1, juvenile interactions2, opposite and same sex interactions345, and territorial interactions6. For decades, the USVs have been used by investigators as proxies to study neuropsychiatric and developmental or behavioral disorders789, and more recently to understand mechanisms and evolution of vocal communication among vertebrates10. Within the sexual interactions, adult male mice produce USV songs, which have some features similar to courtship songs of songbirds11. The use of such multisyllabic repertoires can increase potential flexibility and information they carry, as they can be varied in how elements are organized and recombined, namely syntax. In this protocol a reliable method to elicit USV songs from male mice in various social contexts, such as exposure to fresh female urine, anesthetized animals, and estrus females is described. This includes conditions to induce a large amount of syllables from the mice. We reduce recording of ambient noises with inexpensive sound chambers, and present a quantification method to automatically detect, classify and analyze the USVs. The latter includes evaluation of call-rate, vocal repertoire, acoustic parameters, and syntax. Various approaches and insight on using playbacks to study an animal's preference for specific song types are described. These methods were used to describe acoustic and syntax changes across different contexts in male mice, and song preferences in female mice.

High‐throughput analysis of vocalizations reveals sex‐specific changes in Fmr1 mutant pups

“…However, future studies should further compare the different types of analyses and perhaps include a hand-scored analysis of the Holy and Guo taxonomy to improve understanding of the capacity of the proposed automated analysis to accurately extract data. 24 Secondly, the use of both systems may be advantageous in identifying the changes following loss of Fmr1. Previous studies have indicated that declines in body temperature can map onto increases in the number of vocalizations produced.…”

“…24 In order to determine whether the automated method would be suitable for neonatal vocalizations, we measured syllable production, as well as acoustic and temporal properties, then compared the results to the results obtained using previously described methodology. 24 In order to determine whether the automated method would be suitable for neonatal vocalizations, we measured syllable production, as well as acoustic and temporal properties, then compared the results to the results obtained using previously described methodology.…”

“…24 In the Sonogram Parameters section, we set Min Frequency to 15,000 Hz, Max Frequency to 125,000 Hz, the sampling frequency to 256 kHz and the Threshold to 0.3. Sonograms were then processed using the graphical user interface as described previously.…”

“…[24][25][26][27] For other forms of behavioral analysis, large efforts have been made to automate data collection, such as video tracking in Caenorhabditis elegans models 28 or gesture tracking in mice. Conducting vocalization analyses is timeconsuming and often subject to software malfunction, both of which may contribute to the paucity of studies.…”

Recurrent laryngeal nerve transection in mice results in translational upper airway dysfunction

“…Ultrasonic vocalization testing was performed to assess vocal function in mice 1 week prior to surgery and at 1, 2, and 5 WPS. Two to three nights prior to vocalization testing, mice were co-housed with a sexually mature female mouse overnight to sexually experience the male mice, as sexually experienced male mice are likely to produce greater numbers of ultrasonic calls (Chabout, Jones-Macopson, & Jarvis, 2017 (Grant et al, 2014;Grant et al, 2015;Hammerschmidt et al, 2012). Offline acoustic analysis of vocal recordings for each mouse were analyzed using SASLab Pro (Avisoft, Germany).…”