Attraction of female house mice to male ultrasonic courtship vocalizations depends on their social experience and estrous stage

PLOS ONE RESEARCH ARTICLE Attraction of female house mice to male ultrasonic courtship vocalizations depends on their social experience and estrous stage Jakob Beck ID, Bettina Wernisch, Teresa Klaus, Dustin J. Penn*, Sarah M. Zala Department of Interdisciplinary Life Sciences, Konrad Lorenz Institute of Ethology, University of Veterinary Medicine Vienna, Vienna, Austria * a1111111111 a1111111111 a1111111111 a1111111111 a1111111111 OPEN ACCESS Citation: Beck J, Wernisch B, Klaus T, Penn DJ, Zala SM (2023) Attraction of female house mice to male ultrasonic courtship vocalizations depends on their social experience and estrous stage. PLoS ONE 18(10): e0285642. https://doi.org/10.1371/ journal.pone.0285642 Editor: Dragan Hrncic, Belgrade University Faculty of Medicine, SERBIA Received: April 25, 2023 Accepted: September 25, 2023 Published: October 10, 2023 Copyright: © 2023 Beck et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Abstract Male house mice (Mus musculus) produce complex ultrasonic vocalizations (USVs), especially during courtship and mating. Playback experiments suggest that female attraction towards recordings of male USVs depends on their social experience, paternal exposure, and estrous stage. We conducted a playback experiment with wild-derived female house mice (M. musculus musculus) and compared their attraction to male USVs versus the same recording without USVs (background noise). We tested whether female attraction to USVs is influenced by the following factors: (1) social housing (two versus one female per cage); (2) neonatal paternal exposure (rearing females with versus without father); and (3) estrous stage. We found that females showed a significant attraction to male USVs but only when they were housed socially with another female. Individually housed females showed the opposite response. We found no evidence that pre-weaning exposure to a father influenced females’ preferences, whereas estrous stage influenced females’ attraction to male USVs: females not in estrus showed preferences towards male USVs, whereas estrous females did not. Finally, we found that individually housed females were more likely to be in sexually receptive estrous stages than those housed socially, and that attraction to male USVs was most pronounced amongst non-receptive females that were socially housed. Our findings indicate that the attraction of female mice to male USVs depends upon their social experience and estrous stage, though not paternal exposure. They contribute to the growing number of studies showing that social housing and estrous stage can influence the behavior of house mice and we show how such unreported variables can contribute to the replication crisis. Data Availability Statement: All relevant data are within the paper and its Supporting Information files. Funding: Our research was supported by Austrian Science Fund (FWF P28141-B25 and P36446-B) (http://www.fwf.ac.at) and Human Frontier Science Program (RGP0003/2020) to DJP and SMZ. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Introduction House mice (Mus musculus) emit ultrasonic vocalizations (USVs) during courtship and mating, and the number of USVs emitted during sexual interactions correlates with males’ mating and reproductive success [1,2]. USVs have been described in wild-derived mice [3,4] as well as laboratory mice [5–9] in a variety of contexts. They are emitted as discrete sounds ("calls") that PLOS ONE | https://doi.org/10.1371/journal.pone.0285642 October 10, 2023 1 / 22 PLOS ONE Competing interests: The authors have declared that no competing interests exist. Attraction of female mice to male ultrasonic courtship vocalizations can be automatically detected [10] and classified into several different classes (at least 3 to 12 "syllable" types), according to their shape, complexity, and other spectrographic features [11]. Male courtship USVs are uttered in repeated phrases that vary in syllable sequences ("syntax") and several other features comparable to the songs of birds and cetaceans [9]. Both sexes emit USVs during opposite-sex interactions [12,13] and their calls share similar qualitative features [13], though males produce most calls (ca. 85% to 93%) during opposite-sex interactions [13,14]. Male USVs are individually distinctive [15,16] and differ among laboratory strains [17]. Yet, males also modulate the emission of courtship USVs depending upon several factors, including their health [18], social housing [19], socio-sexual experience [20–22], sex of the stimulus individual [13], and estrous stage of a stimulus female [23]. Females’ responses towards male USVs have been investigated using playback experiments [2,4,24–29]. Females exhibit greater vocal responses to USVs than do males [26], and they are more attracted to complex than simple types of male USVs [25]. Females show more attraction to the USVs of males of their own species than towards an unfamiliar Mus species [27] and towards USVs of unfamiliar non-kin versus familiar male siblings [4]. The aim of our present study was to investigate factors proposed to influence female attraction to male USVs. Three different factors have been suggested to influence female attraction to male USVs (which are similar to factors proposed to influence female attraction to male scent [30,31]): Firstly, habituation responses of female (CBA) mice to male USVs were influenced by whether they were kept in individual housing (IH) versus social housing (SH) [32]. Housing did not influence female attraction to USVs versus white noise, but IH mice and SH (with females) surprisingly showed increased attraction over time, whereas SH mice (with males and females) showed no increase over time. Social housing has been found to influence several behaviors in mice and other rodents [33], including sexual behaviors and auditory mechanisms that could alter female responses to USVs. For example, male mice [34] and rats (Rattus norvegicus) [35] kept in IH show lower sexual motivation than SH males, and female mice socially isolated during puberty exhibit less receptive sexual behaviors (lordosis) compared to socially reared individuals [36]. Mice kept in IH versus SH show differences in auditory perception [37], including perception of USVs [38], and neural auditory mechanisms [39–41]. These findings are interesting, but also concerning because studies on laboratory rodents rarely provide information about social housing conditions, and such unreported variables potentially contribute to the replication crisis [42–45]. Secondly, another playback study found that female attraction to USVs of males from a different strain depended on whether females had been reared with th (...truncated)