LMT USV Toolbox, a Novel Methodological Approach to Place Mouse Ultrasonic Vocalizations in Their Behavioral Contexts—A Study in Female and Male C57BL/6J Mice and in Shank3 Mutant Females

Abstract

Ultrasonic vocalizations (USVs) are used as a phenotypic marker in mouse models of neuropsychiatric disorders. Nevertheless, current methodologies still require time-consuming manual input or sound recordings clean of any background noise. We developed a method to overcome these two restraints to boost knowledge on mouse USVs. The methods are freely available and the USV analysis runs online at https://usv.pasteur.cloud. As little is currently known about usage and structure of ultrasonic vocalizations during social interactions over the long-term and in unconstrained context, we investigated mouse spontaneous communication by coupling the analysis of USVs with automatic labeling of behaviors. We continuously recorded during 3 days undisturbed interactions of same-sex pairs of C57BL/6J sexually naive males and females at 5 weeks and 3 and 7 months of age. In same-sex interactions, we observed robust differences between males and females in the amount of USVs produced, in the acoustic structure and in the contexts of emission. The context-specific acoustic variations emerged with increasing age. The emission of USVs also reflected a high level of excitement during social interactions. We finally highlighted the importance of studying long-term spontaneous communication by investigating female mice lacking Shank3, a synaptic protein associated with autism. While the previous short-time constrained investigations could not detect USV emission abnormalities, our analysis revealed robust differences in the usage and structure of the USVs emitted by mutant mice compared to wild-type female pairs.

Transient reduction in dendritic spine density in brain-specific profilin1 mutant mice is associated with behavioral deficits

“…Instead, the number of ultrasonic vocalizations (USV) was affected. USV is mostly emitted during active social behaviors in mice and is a prominent aspect of their social behavior repertoire ( Sungur et al, 2018 ; de Chaumont et al, 2021 ). They serve important communicative functions and are typically altered in mouse models for neurodevelopmental disorders ( Caruso et al, 2020 ).…”

From Mating to Milk Access: A Review of Reproductive Vocal Communication in Mice

“…So, although there is published data on laboratory and wild-derived mice, no studies were performed in naturalistic conditions. Nevertheless, efforts have been made to surpass this, and de Chaumont et al (2021) have recently developed a sophisticated apparatus that allows the recording of video and sounds during long periods of time which can provide novel insights into vocal communication and its correlation with behaviour.…”

“…As such, it would be expected that females communicate vocally in these contexts and that this would have an impact on maternal care and pup survival. But the current literature only addresses vocal communication in same sex or heterosexual encounters ( Warren et al, 2018 , 2020 ; Sasaki et al, 2020 ; de Chaumont et al, 2021 ). It is know that females vocalise in same-sex encounters and produce more complex USVs in these situations ( Matsumoto and Okanoya, 2018 ) but it is still unknown how this would be affected by the presence of a male.…”

Ultrasonic Vocalizations in Adult C57BL/6J Mice: The Role of Sex Differences and Repeated Testing

“…Several studies have tried to analyze differences in USVs between male and female mice (Hammerschmidt et al, 2012 ; von Merten et al, 2014 ; Zala et al, 2017 ; Matsumoto and Okanoya, 2018 ; de Chaumont et al, 2021 ), also with novel technical approaches such as deep learning networks (Ivanenko et al, 2020 ), yielding to the emergence of a variety of either quantitative or qualitative differences (or both) without, so far, a univocal pattern. Divergences among these studies mainly arise from differences in testing procedures, e.g., sex of the stimulus animal and pre-testing isolation conditions.…”

“…Divergences among these studies mainly arise from differences in testing procedures, e.g., sex of the stimulus animal and pre-testing isolation conditions. The sex of the “receiver” is known to critically modulate several characteristics of USVs of the “emitter” in mice of both sexes (Zala et al, 2017 ); for instance, quantitative sex differences in USVs were described between female-female and male-female interactions (von Merten et al, 2014 ), while both quantitative and qualitative differences were observed in same-sex interactions (de Chaumont et al, 2021 ). Also, most of the studies on female USVs have applied relatively long periods (more than 24 h) of pre-testing isolation in order to induce a resident status in the subject and assure the identification of the emitter (Moles et al, 2007 ; Hammerschmidt et al, 2012 ).…”