Female mice in established social groups use different ultrasonic vocalizations during peaceful and aggressive interactions.

Event Abstract Back to Event Females influence male mouse ultrasonic courtship vocalizations Jessica L. Hanson1* and Laura M. Hurley1 1 Indiana University, United States The laboratory mouse is an emerging model for context-dependent vocal signaling and reception. Mouse ultrasonic vocalizations (USVs) are produced primarily in social contexts, most intensely by males courting females. The presence of a female will trigger male calls, but these vocalizations can be induced by presentation of female urine; and female mice respond to these vocalizations with approach behavior. USVs are complex, occurring in distinct syllables that can be grouped into several types and are produced across species and strains of mice. Parameters of these syllables, such as duration and frequency (kHz) are unique to individuals, vary across development, and depend on social housing conditions. The behavioral significance of different syllable types, including the contexts in which different vocalizations are made, is not well-understood, nor is the response of female mice to different types of USVs. Here, we examined the influence of female presence and estrous state on male courtship USVs by allowing males to interact with females of different estrous states, and subsequently removing the females while continuing to record male USVs. We also collected USV and behavioral data from courtship interactions while females were tethered by a headstage (cannulation for another study). In these two experiments we explored relationships between vocalizations and other courtship behaviors, including correlating percent behavior per trial as well assessing temporal relationships between behaviors throughout trials. These experimental manipulations produced three main findings: 1) the frequency (kHz), bandwidth, and duration of syllables produced by males were influenced by the estrous phase of female partners, 2) syllable types changed when females were removed, and 3) vocal behavior was related to other courtship behaviors. These findings show that mouse ultrasonic vocalizations are sensitive to changes in female phase and presence, and provide support for the idea that courtship USVs have the potential to communicate information about the behavioral context of the male. Keywords: Courtship, female state, Mouse, ultrsonic vocalization Conference: Tenth International Congress of Neuroethology, College Park. Maryland USA, United States, 5 Aug - 10 Aug, 2012. Presentation Type: Poster (but consider for student poster award) Topic: Communication Citation: Hanson JL and Hurley LM (2012). Females influence male mouse ultrasonic courtship vocalizations. Conference Abstract: Tenth International Congress of Neuroethology. doi: 10.3389/conf.fnbeh.2012.27.00255 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 30 Apr 2012; Published Online: 07 Jul 2012. * Correspondence: Ms. Jessica L Hanson, Indiana University, Bloomington, Indiana, United States, jeshanso@indiana.edu Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Jessica L Hanson Laura M Hurley Google Jessica L Hanson Laura M Hurley Google Scholar Jessica L Hanson Laura M Hurley PubMed Jessica L Hanson Laura M Hurley Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

We examined the individual context of ultrasonic vocalizations (USVs) produced by free-living wild male and female adult brush mice (Peromyscus boylii). We tested the hypothesis that USV production is dependent on behavioral context, and is important during both adult male and female interactions. Our methods included a 12-channel microphone array, radio-telemetry and thermal imaging that allowed us to determine: (1) who produced USVs, (2) characteristics of USVs, (3) type of USVs, (4) behavioral context of USVs and (5) the identity of the second mouse if an individual was not alone when a USV was produced. Females vocalized as much as males and produced the same types of USVs as males. There were no differences between spectral characteristics of male and female USVs. Females and males vocalized in the presence of one another. Importantly, when females were together they vocalized more than expected based on the proportion of time they spent together. Our results suggest that, in addition to facilitating courtship and mating, USVs are general territorial calls for neighbors because females vocalized in the presence of their neighbors. Despite a large literature on laboratory mouse (Mus musculus) USVs, studies are heavily biased towards males. Our results on brush mice, a species with a similar breeding system to the lab mouse and other rodents, suggest that female–female communication is an important and underappreciated component of the evolution and maintenance of mouse USVs.

Mice produce ultrasonic vocalizations (USVs) in a wide variety of social contexts, including courtship, investigation, and territorial defense. Despite the belief that mouse USVs are innate, social experience may be necessary for mice to learn the appropriate situation to emit USVs. Mouse USVs have been divided into categories based on their spectrotemporal parameters, but it is currently unclear if social experience changes these parameters (e.g., frequency and duration) or the proportion of calls from each category produced. Social isolation has been found to influence USV production in male mice. To investigate the influence of social isolation on vocal behavior in female mice, recordings were made of USVs emitted to unfamiliar male and female mice by subjects with one of three types of social experience. Twenty-four adult female CBA/CaJ mice either lived alone, lived with other females only, or lived with other females and had limited access to a male. Mice were recorded while in isolation, ensuring all recorded USVs were from the female of interest. Vocalizations were separated into nine categories and peak frequency, duration, and bandwidth were measured for every call. Socially isolated mice did not produce significantly more USVs or USV types than socially experienced mice. Social isolation did not have a significant effect on the features of USVs, suggesting production of USVs may not be learned in female mice.

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.

Ultrasonic vocalizations (USVs) in rodents can be used as a method of measuring the affective state of the animal during a wide range of experiences. Adult rats generally emit two types of USVs: “alarm” calls (22kHz) and calls during “nonaggressive” behavioral situations (50kHz). Rat USVs can therefore be measured and classified when a rat is in the presence of a natural predator. In humans, PTSD results from exposure to a traumatic event(s) which evoke a level of fear, helplessness, or horror with a threat of physical harm. In this study, a rodent model of traumatic stress was used to elicit a traumatic event via (a single protected) exposure to a predator species (snake, ferret, or cat). To determine the impact of these animals as a natural predator species, we examined each one individually and assessed USVs to determine the impact on ultrasonic communication by these predator species. Rats (~11–12 weeks old) were behaviorally assessed pre‐ and post‐predator exposure on several behavioral tasks. USVs were recorded during the open field task and on predator exposure days (pre‐ and post‐exposure, in transfer cages, and during predator exposure, in the predator home cage). To record the USVs, an Avisoft‐Ultra SoundGate electret 116Hnb microphone and Recorder USGH program (Avisoft Bioacoutistc, Germany) were used. Rats were placed in a protective predator tube, and then placed in the predator home cage. These tubes allow for visual and olfactory cues; however, prevent the rat from gross physical contact. The rats were exposed to one of the predators or sham (using a predator tube free of predator scent). Sham‐exposed rats emitted significantly more 50kHz USVs than any of the predator‐ exposed rats (F(3,44)= 9.12, p < .0001). Cat‐exposed rats did not emit any 50kHz USVs during exposure, but emitted the most 22kHz USVs, significantly more than any other predator or sham (F(3,44)= 3.29, p < .05). 22kHz USVs were emitted by 75% of cat‐exposed rats and only 2 of 12 snake‐exposed rats. Rats exposed to the ferret and sham did not emit any fear‐induced 22kHz vocalizations, only 50kHz vocalizations. Based on emitted ultrasonic calls, the cat appears to be the most impactful, eliciting the most fear‐induced vocalizations. To the contrary, the ferret did not cause any fear‐inducing vocalizations. This study has provided the ground work for the recording of USVs during our laboratory's rodent model of traumatic stress. For future research, this method can be used as an additional measure, along with assessing overt behavior measures, to identify candidate therapeutic compounds for the prevention and/or treatment of PTSD.Support or Funding InformationMaterial has been reviewed by the Walter Reed Army Institute of Research. There is no objection to its presentation and/or publication. The opinions or assertions contained herein are the private views of the author, and are not to be construed as official, or as reflecting true views of the Department of the Army or the Department of Defense. Research was conducted in an AAALACi accredited facility in compliance with the Animal Welfare Act and other federal statutes and regulations relating to animals and experiments involving animals and adheres to principles stated in the Guide for the Care and Use of Laboratory Animals, NRC Publication, 2011 edition. Supported by the Military Operational Medicine Research Program, US Army Medical Research and Materiel Command.

Spectrographic analyses reveal signals of individuality and kinship in the ultrasonic courtship vocalizations of wild house mice

The effects of aging and sex on detection of ultrasonic vocalizations by adult CBA/CaJ mice (Mus musculus)

Although ultrasonic vocalizations (USVs) have been recorded in many species of rodent and in various contexts, e.g. sexual behaviour and aggression, it has not been demonstrated for the endangered Turkish Spiny Mouse Acomys cilicius Spitzenberger. This study investigated whether A. cilicius emits USVs and, if so, how these USVs associated with non-vocalization behaviour. Ultrasonic recording equipment was set up for 12 days in an off-exhibit enclosure of A. cilicius at Bristol Zoo. At least seven different types of USV were recorded. For eight of the 12 study days, ultrasonic and video recording equipment were run concurrently. From these observations it was found that emission of USVs were associated with sexual behaviour, aggression and social investigation. The results of this study show for the first time that captive A. cilicius produce USVs that resemble those produced by other rodent species, including its close relative the Egyptian Spiny Mouse A. cahirinus Desmarest. As these findings apply only to a captive Turkish Spiny Mouse population, additional work should be carried out to investigate the behaviour and USV production in the wild in addition to further research on captive populations investigating the apparent communicative function of these vocalizations.

Adult mouse ultrasonic vocalizations (USVs) occur in multiple behavioral and stimulus contexts associated with various levels of arousal, emotion and social interaction. Here, in three experiments of increasing stimulus intensity (water; female urine; male interacting with adult female), we tested the hypothesis that USVs of adult males express the strength of arousal and emotion via different USV parameters (18 parameters analyzed). Furthermore, we analyzed two mouse lines with heterozygous Foxp2 mutations (R552H missense, S321X nonsense), known to produce severe speech and language disorders in humans. These experiments allowed us to test whether intact Foxp2 function is necessary for developing full adult USV repertoires, and whether mutations of this gene influence instinctive vocal expressions based on arousal and emotion. The results suggest that USV calling rate characterizes the arousal level, while sound pressure and spectrotemporal call complexity (overtones/harmonics, type of frequency jumps) may provide indices of levels of positive emotion. The presence of Foxp2 mutations did not qualitatively affect the USVs; all USV types that were found in wild-type animals also occurred in heterozygous mutants. However, mice with Foxp2 mutations displayed quantitative differences in USVs as compared to wild-types, and these changes were context dependent. Compared to wild-type animals, heterozygous mutants emitted mainly longer and louder USVs at higher minimum frequencies with a higher occurrence rate of overtones/harmonics and complex frequency jump types. We discuss possible hypotheses about Foxp2 influence on emotional vocal expressions, which can be investigated in future experiments using selective knockdown of Foxp2 in specific brain circuits.

The effect of playback of 22-kHz and 50-kHz ultrasonic vocalizations on rat behaviors assessed with a modified open-field test

Two experiments were conducted to examine 70-kHz ultrasonic courtship vocalizations by adult male mice (Mus musculus) to novel odors following exposure to these odors in infancy and/or adulthood. The research was performed in hopes of better understanding the experiences giving rise to vocalizations to naturally occurring chemosignals. Experiment 1 demonstrated that adult males normally do not vocalize to the urine of female rats but would come to do so if adult female mice odorized with female rat urine were repeatedly encountered postpubertally. On the other hand, encountering their own mother odorized with female rat urine from birth until weaning did not promote vocalizations to the urine of female rats. Experiment 2 was designed to examine vocalizations to the urine of female mice whose urinary odor was altered by the ingestion of fenugreek, a spice. Although the magnitude of the effect was smaller in this experiment, greater amounts of vocalization again were seen by males that as adults encountered females that had ingested fenugreek. Males with such experience also showed a small but significant elevation in vocalizations to the fenugreek odor itself. Again, experience with the novel odor during infancy was not associated with elevated vocalizations during adulthood either to fenugreek-altered urine or to the fenugreek odor itself. Thus vocalizations to two different novel odors occurred only after an adult male had encountered an adult female odorized with the novel odor. On the other hand, none of the novel odor experiences eliminated vocalizations to the naturally occurring chemosignal in female mouse urine.(ABSTRACT TRUNCATED AT 250 WORDS)

Exposure to males or male urinary scent can induce and accelerate the rate of female estrous cycling in house mice ("Whitten effect"), and this response has been replicated many times since its discovery over 60 years ago. Here, we tested whether exposing female mice to recordings of male courtship ultrasonic vocalizations (USVs) induces estrous cycling, and whether exposure to both male scent and USVs has a stronger effect than to either of these stimuli alone. We conducted our study with 60 wild-derived female house mice (Mus musculus musculus). After singly housing females for 14 days, we monitored estrous stages via vaginal cytology for two weeks while isolated from males or male stimuli. We continued monitoring estrus for two more weeks during experimental exposure to one of four different types of stimuli: (1) clean bedding and background noise playback (negative control); (2) recordings of male USVs (16 min per day) and clean bedding (male USV treatment); (3) soiled male bedding and background noise playback (male odor treatment; positive control); or (4) male USVs and soiled male bedding (male odor and USV treatment). Females were then paired with males to test whether any of the four treatments influenced female reproduction (especially latency to birth). We confirmed that exposure to male odor increased female cycling, as expected, but exposure to recordings of male USVs had no effect on estrus. Females exposed to both USVs and odor went through more cycles compared to controls, but did not differ significantly from exposure to male odor (and background noise). After pairing females with a male, females showing male odor-induced cycling produced their first litter sooner than controls, whereas USVs did not have such an effect. This is the first study to our knowledge to show that male odor induces estrus in wild house mice and to show functional effects on reproduction. Our results do not support the hypothesis that male vocalizations induce female estrus, although we suggest other approaches that could be used to further test this hypothesis.

Ultrasonic vocalizations (USV) in rats may communicate "affective" states during pain, sex and aggression. This proposal was evaluated in an experiment with adult male Long-Evans rats during agonistic encounters; specifically, morphine and naltrexone effects were studied on different types of USV by intruder rats exposed to resident attacks and to "threat of attacks" (i.e., intruder residing within the home cage of the resident but prevented from physical contact by a wire mesh cage). Intruders readily emitted USV during agonistic encounters. These calls consisted primarily of two distinct distributions of pure tone whistles: 0.3-3 s, 19-32 kHz ("low") calls and 0.02-0.3 s, 32-64 kHz ("high") calls. Sonographic analysis revealed a considerable repertoire of frequency modulated calls. Different types of vocalizations proved to be differentially sensitive to the opiate treatments: morphine (1-10 mg/kg SC) dose-dependently decreased the rate, duration and pitch of both low and high frequency USV during the threat of attack; this decrease in rate and duration measures was naltrexone-reversible (0.1 mg/kg IP). Interestingly, audible vocalizations were also emitted but were unaffected by morphine in this dose range. Concomitant with the decrease in USV after morphine was a dose-dependent decrease in rearing, walking and nasal contact behavior with increases in submissive crouch behavior and tail flick analgesia. The decreases in rate and duration of both low and high USV and the pitch of specific frequency modulated calls after morphine administration may reflect an attenuation of affective aspects of pain, and the many characteristics of US (rate, duration, pitch, frequency modulation, pre-and suffix attributes and temporal structure) point to potentially diverse functions.(ABSTRACT TRUNCATED AT 250 WORDS)

In rodents, ultrasonic vocalizations (USVs) are used for communication, and they are widely observed in both wild and laboratory strains. Several types of USVs are commonly found in mice and rats. Here, we review these USVs, discuss the contexts in which mice emit USVs, and compare the characteristics of mouse and rat USVs. Based on their acoustic properties, we propose that USVs function as emotional expressions.

Mice are frequently used as an animal model for human hearing research, yet their hearing capabilities have not been fully explored. Previous studies (Henry, 2004; Radziwon et al., 2009) have established auditory threshold sensitivities for pure tone stimuli in CBA/CaJ mice using ABR and behavioral methodologies. Yet, little is known about how they perceive their own ultrasonic vocalizations (USVs), and nothing is known about how aging influences this perception. The aim of the present study was to establish auditory threshold sensitivity for several types of USVs, as well as to track these thresholds across the mouse’s lifespan. In order to determine how well mice perceive these complex communication stimuli, several CBA/CaJ mice were trained and tested at various ages in a detection task using operant conditioning procedures. Results showed that mice were able to detect USVs well into their lifespan, and that thresholds differed across USV types. Male mice showed higher thresholds for certain USVs later in life than females. In conclusion, the results suggest that mice are sensitive to their complex vocalizations even into old age, highlighting their likely importance for survival and communication.