Female Ultrasonic Vocalizations Research Articles

Classifying sex and strain from mouse ultrasonic vocalizations using deep learning.

Vocalizations are widely used for communication between animals. Mice use a large repertoire of ultrasonic vocalizations (USVs) in different social contexts. During social interaction recognizing the partner's sex is important, however, previous research remained inconclusive whether individual USVs contain this information. Using deep neural networks (DNNs) to classify the sex of the emitting mouse from the spectrogram we obtain unprecedented performance (77%, vs. SVM: 56%, Regression: 51%). Performance was even higher (85%) if the DNN could also use each mouse's individual properties during training, which may, however, be of limited practical value. Splitting estimation into two DNNs and using 24 extracted features per USV, spectrogram-to-features and features-to-sex (60%) failed to reach single-step performance. Extending the features by each USVs spectral line, frequency and time marginal in a semi-convolutional DNN resulted in a performance mid-way (64%). Analyzing the network structure suggests an increase in sparsity of activation and correlation with sex, specifically in the fully-connected layers. A detailed analysis of the USV structure, reveals a subset of male vocalizations characterized by a few acoustic features, while the majority of sex differences appear to rely on a complex combination of many features. The same network architecture was also able to achieve above-chance classification for cortexless mice, which were considered indistinguishable before. In summary, spectrotemporal differences between male and female USVs allow at least their partial classification, which enables sexual recognition between mice and automated attribution of USVs during analysis of social interactions.

Intranasal administration of exosomes derived from mesenchymal stem cells ameliorates autistic-like behaviors of BTBR mice

Autism spectrum disorders (ASD) are neurodevelopmental disorders characterized by three core symptoms that include social interaction deficits, cognitive inflexibility, and communication disorders. They have been steadily increasing in children over the past several years, with no effective treatment. BTBR T+tf/J (BTBR) mice are an accepted model of evaluating autistic-like behaviors as they present all core symptoms of ASD. We have previously shown that transplantation of human bone marrow mesenchymal stem cells (MSC) to the lateral ventricles of BTBR mice results in long lasting improvement in their autistic behavioral phenotypes. Recent studies point exosomes as the main mediators of the therapeutic effect of MSC. Here, we tested whether treatment with the exosomes secreted from MSC (MSC-exo) will show similar beneficial effects. We found that intranasal administration of MSC-exo increased male to male social interaction and reduced repetitive behaviors. Moreover, the treatment led to increases of male to female ultrasonic vocalizations and significant improvement in maternal behaviors of pup retrieval. No negative symptoms were detected following MSC-exo intranasal treatments in BTBR or healthy C57BL mice. The marked beneficial effects of the exosomes in BTBR mice may translate to a novel, non-invasive, and therapeutic strategy to reduce the symptoms of ASD.

Ultrasonic vocalizations of female Norway rats (Rattus norvegicus) in response to social partners.

In many species of animals, male vocalizations function to attract mating partners and coordinate sexual interactions. Whereas male vocalizations have been well studied in several species, the function of female vocalizations in mating contexts is not fully understood. In Norway rats (Rattus norvegicus), both males and females produce ultrasonic vocalizations (USVs) during sexual encounters with opposite sex partners. The aim of this study was to test the hypothesis that female vocalizations play a role in sociosexual interactions by examining how rates of 50 kHz USV production vary in relation to the sex and gonadal status of the partner, and by examining whether the proportion of frequency modulated (FM) and constant frequency calls differs between these categories of social partner. The results showed that females produced a higher total number of 50 kHz USVs to intact males than castrated males, and produced similar numbers of calls to both categories of females. Females also produced a higher proportion of FM calls to male partners than to female partners, and spent more time in the vicinity of male than female partners, regardless of the partners' gonadal status. Female USVs therefore potentially provide a measure of sexual motivation and may function to promote female mate choice in this species with multimale mating and a high risk of infanticide.

Female ultrasonic vocalizations have no incentive value for male rats.

Ultrasonic vocalizations (USVs) are emitted in response to a sexual partner before, during, and after copulation. These vocalizations are the so-called 50-kHz USVs and can be subdivided into flat and frequency-modulated (FM) 50-kHz trill calls. In the present series of experiments, the potential unconditioned and conditioned incentive properties of female 50-kHz USVs for male rats were examined. USVs were recorded from sexually receptive females during the precopulatory phase. A complete 10-min song, or single flat or FM trill calls were selected as auditory stimuli for Experiments 1 and 2. As FM trill calls, a multistep call was used in Experiments 1 and 2a, and an upward ramp call was used in Experiment 2b. The auditory stimuli were played back with a loudspeaker to naïve and sexually experienced male rats in a sexual incentive motivation test. The odor of a sexually receptive female rat was also used as an incentive stimulus for comparison. In a third experiment, a devocalized female, a sham female and a male rat were used as incentive stimuli. It was found that the auditory stimuli did not induce approach behavior in naïve and sexually experienced male rats, but the olfactory stimulus did. In addition, the males spent equal amounts of time in the vicinity of devocalized and vocalizing females. These data show that 50-kHz USVs neither are unconditioned nor conditioned incentives for male rats.

Female and male adult brush mice (Peromyscus boylii) use ultrasonic vocalizations in the wild

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.

The Structure and Usage of Female and Male Mouse Ultrasonic Vocalizations Reveal only Minor Differences

Ultrasonic vocalizations (USV) of mice are increasingly recognized as informative dependent variables in studies using mouse models of human diseases. While pup vocalizations primarily serve to re-establish contact with the mother, adult male “songs” were considered to be courtship signals. Alternatively, mouse USVs may generally function as territorial signals. To distinguish between these two hypotheses, we compared the structure and usage of adult male and female USVs in staged resident-intruder encounters. If calls function primarily as courtship signals, males should respond stronger than females, specifically when presented with a female intruder. Refuting this hypothesis, we found that in response to female intruders, females called more than males (228±32 calls/min vs. 71±15 calls/min), and males called more to female than to male intruders (14±7.5 calls/min). There were no significant differences in the acoustic characteristics of the calls given by females and males. To control for the influence of the intruder's behavior on calling, we repeated the experiments using anaesthetized intruders. Again, females produced more calls to female than male intruders (173±17 calls/min vs. 71±15 calls/min), while males called more in response to female than male intruders (39±17 calls/min), and there were no acoustic differences in female and male calls. The vocal activity did not differ significantly with regard to intruder state (awake or anaesthetized), while the acoustic structure exhibited significant differences. Taken together, our findings support the view that calls do not mainly function as courtship signals, although they might serve both a territorial (sex-independent) and a courtship function. The comparison of responses to awake vs. anaesthetized intruders suggests that the latter are sufficient to elicit vocal activity. The subtle acoustic differences, however, indicate that the subject differentiates between intruder states.

Lack of social exploratory activation in male μ-opioid receptor KO mice in response to playback of female ultrasonic vocalizations

The opioid system controls social behavior and it was hypothesized that it therefore plays a role in neuropsychiatric disorders such as autism that are characterized by social and communication deficits. Mice communicate via ultrasonic vocalizations. As pups, they produce ultrasonic vocalizations when isolated from dam and littermates. These calls serve an important biological purpose, since they elicit search and retrieval behavior in the mother. Administration of μ-opioid-receptor-agonists diminishes such isolation-induced ultrasonic vocalizations and μ-opioid-receptor knock-out mouse pups (Orpm –/–) emit fewer ultrasonic vocalizations during isolation than intact controls (Orpm +/+). In adulthood, male and female mice produce ultrasonic vocalizations during social interactions. However, little is known about occurrence and function of ultrasonic vocalizations produced by adult females. Here, we conducted a playback experiment in order to assess whether female ultrasonic vocalizations elicit changes in the recipient's behavior and whether a possible change in behavior is dependent on a functioning opioid system by comparing Orpm –/– mice with Orpm +/+ controls. Our results showed that female ultrasonic vocalizations elicit exploratory activity in male recipients and that elicitation of exploratory activity in response to female ultrasonic vocalizations is dependent on an intact opioid system, since such a response was not seen in Orpm –/– mice. Lack of exploratory activation seen in Orpm –/– mice is unlikely due to hearing deficits as shown by an auditory cued fear-conditioning-task. Hence, these findings support the phenotypic relevance of Orpm –/– mice for the study of autism.