Ultrasonic Calls Research Articles

Field tests reveal acoustic variation of call types in a subterranean rodent, the Northern Mole Vole Ellobius talpinus

Abstract This study investigates acoustic variation of human-audible sonic (below 20 kHz) and human-inaudible ultrasonic (above 20 kHz) calls in a wild subterranean rodent, the Northern Mole Vole (Ellobius talpinus), under 3 call-eliciting tests conducted during captures for 1 day. The Contact-in-Tunnel Test modeled contacts of 2 individuals during digging earth in a burrow tunnel. The Restraint Test modeled restraint of an animal by a surrogate predator (human hand). The Release-to-Burrow Test modeled acoustic communication of many family members returned to their home burrow after their isolation for about 8 h, from morning to evening. We described 8 call types: 3 sonic, 3 ultrasonic, and 2 expanding from sonic to ultrasonic range of frequencies; 6 call types were described for the first time for this species. No relationship was found between acoustic parameters and proxies of body size (body mass and the width of 2 upper incisors). No sex differences were found in body size or the acoustic parameters. Different call types prevailed in different tests: wheeks and upsweeps were made during peaceful interactions; squeaks and squeals were related to animal discomfort during the Restraint Test; rasps were only made in Release-to-Burrow Tests when animals were plugging the burrow entrance; and variative calls did not show any relationship with type of test. Based on presence or absence of certain call types in the tests, we evaluate their potential communicative role in comparison with published data on vocal repertoires of other subterranean rodents.

Early ultrasonic vocalization deficits and related thyroarytenoid muscle pathology in the transgenic TgF344-AD rat model of Alzheimer's disease.

Alzheimer's disease (AD) is a progressive neurologic disease and the most common cause of dementia. Classic pathology in AD is characterized by inflammation, abnormal presence of tau protein, and aggregation of β-amyloid that disrupt normal neuronal function and lead to cell death. Deficits in communication also occur during disease progression and significantly reduce health, well-being, and quality of life. Because clinical diagnosis occurs in the mid-stage of the disease, characterizing the prodrome and early stages in humans is currently challenging. To overcome these challenges, we use the validated TgF344-AD (F344-Tg(Prp-APP, Prp-PS1)19/Rrrc) transgenic rat model that manifests cognitive, behavioral, and neuropathological dysfunction akin to AD in humans. The overarching goal of our work is to test the central hypothesis that pathology and related behavioral deficits such as communication dysfunction in part manifest in the peripheral nervous system and corresponding target tissues already in the early stages. The primary aims of this study are to test the hypotheses that: (1) changes in ultrasonic vocalizations (USV) occur in the prodromal stage at 6 months of age and worsen at 9 months of age, (2) inflammation as well as AD-related pathology can be found in the thyroarytenoid muscle (TA) at 12 months of age (experimental endpoint tissue harvest), and to (3) demonstrate that the TgF344-AD rat model is an appropriate model for preclinical investigations of early AD-related vocal deficits. USVs were collected from male TgF344-AD (N = 19) and wildtype (WT) Fischer-344 rats (N = 19) at 6 months (N = 38; WT: n = 19; TgF344-AD: n = 19) and 9 months of age (N = 18; WT: n = 10; TgF344-AD: n = 8) and acoustically analyzed for duration, mean power, principal frequency, low frequency, high frequency, peak frequency, and call type. RT-qPCR was used to assay peripheral inflammation and AD-related pathology via gene expressions in the TA muscle of male TgF344-AD rats (n = 6) and WT rats (n = 6) at 12 months of age. This study revealed a significant reduction in mean power of ultrasonic calls from 6 to 9 months of age and increased peak frequency levels over time in TgF344-AD rats compared to WT controls. Additionally, significant downregulation of AD-related genes Uqcrc2, Bace2, Serpina3n, and Igf2, as well as downregulation of pro-inflammatory gene Myd88 was found in the TA muscle of TgF344-AD rats at 12 months of age. Our findings demonstrate early and progressive vocal deficits in the TgF344-AD rat model. We further provide evidence of dysregulation of AD-pathology-related genes as well as inflammatory genes in the TA muscles of TgF344-AD rats in the early stage of the disease, confirming this rat model for early-stage investigations of voice deficits and related pathology.

Deep learning algorithms reveal increased social activity in rats at the onset of the dark phase of the light/dark cycle.

The rapid decrease of light intensity is a potent stimulus of rats' activity. The nature of this activity, including the character of social behavior and the composition of concomitant ultrasonic vocalizations (USVs), is unknown. Using deep learning algorithms, this study aimed to examine the social life of rat pairs kept in semi-natural conditions and observed during the transitions between light and dark, as well as between dark and light periods. Over six days, animals were video- and audio-recorded during the transition sessions, each starting 10 minutes before and ending 10 minutes after light change. The videos were used to train and apply the DeepLabCut neural network examining animals' movement in space and time. DeepLabCut data were subjected to the Simple Behavioral Analysis (SimBA) toolkit to build models of 11 distinct social and non-social behaviors. DeepSqueak toolkit was used to examine USVs. Deep learning algorithms revealed lights-off-induced increases in fighting, mounting, crawling, and rearing behaviors, as well as 22-kHz alarm calls and 50-kHz flat and short, but not frequency-modulated calls. In contrast, the lights-on stimulus increased general activity, adjacent lying (huddling), anogenital sniffing, and rearing behaviors. The animals adapted to the housing conditions by showing decreased ultrasonic calls as well as grooming and rearing behaviors, but not fighting. The present study shows a lights-off-induced increase in aggressive behavior but fails to demonstrate an increase in a positive affect defined by hedonic USVs. We further confirm and extend the utility of deep learning algorithms in analyzing rat social behavior and ultrasonic vocalizations.

Pro-social and pro-cognitive effects of LIT-001, a novel oxytocin receptor agonist in a neurodevelopmental model of schizophrenia

Social and cognitive dysfunctions are the most persistent symptoms of schizophrenia. Since oxytocin (OXT) is known to play a role in social functions and modulates cognitive processes, we investigated the effects of a novel, nonpeptide, selective OXT receptor agonist, LIT-001, in a neurodevelopmental model of schizophrenia. Administration of methylazoxymethanol acetate (MAM; 22 mg/kg) on the 17th day of rat pregnancy is known to cause developmental disturbances of the brain, which lead to schizophrenia-like symptomatology in the offspring. Here, we examined the effects of acutely administered LIT-001 (1, 3, and 10 mg/kg) in MAM-exposed males and females on social behaviour, communication and cognition. We report that MAM-treated adult male and female rats displayed reduced social behaviour, ultrasonic communication and novel object recognition test performance. LIT-001 partially reversed these deficits, increasing the total social interaction time and the number of ‘positive’, highly-modulated 50 kHz ultrasonic calls in male rats. The compound ameliorated MAM-induced deficits in object discrimination in both sexes. Present results confirm the pro-social activity of LIT-001 and demonstrate its pro-cognitive effects following acute administration.

Acute anxiogenic effects of escitalopram are associated with mild alterations in D-amphetamine-induced behavior and social approach evoked by playback of 50-kHz ultrasonic vocalizations in rats

Rats communicate through auditory signals in the ultrasonic range, so-called ultrasonic vocalizations (USV). Short, high-frequency 50-kHz USV are associated with positive affective states and are emitted in appetitive situations, often rewarding social interactions, such as rough-and-tumble play and mating. Exaggerated levels of 50-kHz USV emission can be observed in response to psychostimulants, most notably d-amphetamine (AMPH). There is robust evidence suggesting that 50-kHz USV serve as affiliative signals and help to maintain or re-establish social proximity. A key neurotransmitter involved in behavioral regulation is serotonin (5-hydroxytryptamine, 5-HT). This includes both, the regulation of anxiety-related behavior and ultrasonic communication. Here, we show that acute treatment with the selective 5-HT reuptake inhibitor (SSRI) escitalopram (ESC) leads to increased anxiety-related behavior in the elevated plus maze and tested whether such acute anxiogenic effects of ESC result in alterations in ultrasonic communication in sender and/or receiver. To this aim, we conducted a dose-response study in male rats and assessed AMPH-induced hyperactivity and 50-kHz ultrasonic calling in the sender and social approach behavior evoked by playback of pro-social 50-kHz USV in the receiver. Acute ESC treatment affected both, sender and receiver. This was reflected in a lack of AMPH-induced changes in acoustic features of 50-kHz USV and absence of social exploratory behavior evoked by 50-kHz USV playback, respectively. Albeit the SSRI effects were relatively mild, this supports the notion that the 5-HT system is involved in the regulation of a key aspect of the social behavior repertoire of rodents, namely socio-affective communication through 50-kHz USV.

The Difference between Male and Female Rats in Terms of Freezing and Aversive Ultrasonic Vocalization in an Active Avoidance Test

We examined sex differences in behavioral responses to fear-related learning through ultrasonic vocalization. Adult male and female rats of two inbred rat strains (WAG/Rij rats with genetic predisposition absence epilepsy, n = 25; and the minor substrain NEW with non-epileptic phenotype, n = 32) were examined in the two-way active avoidance task (50 trials). Three behavioral responses were defined: successful learning, unsuccessful learning, and freezing (motionless behavior). We found that males were more likely to experience freezing (40%) than females (3.7%). Females performed significantly better than males: 93% of females showed “successful learning”, while only 43.2% of males did. Long-lasting 22 kHz ultrasonic calls, so-called aversive ultrasonic vocalizations aUSVs, were recorded in 26% of females (only successful learners) and in 50% of males. The density of aUSVs in females was lower than in males. WAG/Rij males performed much poorer than the NEW males. In general, males, in contrast to females, showed significantly poorer avoidance learning, more frequently experienced freezing, and produced more aversive USVs. Males thus experienced a negative emotional state, rather than exhibiting poor cognitive abilities. Perhaps, WAG/Rij rats and a minor NEW substrain showed an increased emotionally in fear-related tasks, because they are prone to emotional disorders or emotional exhaustion. Sex differences in freezing and in aUSVs might result from the poorly understood neuromodulatory effects of the cholinergic system.

Comparison of ultrasonic isolation calls of pure-breeding and interspecies hybrid Phodopus dwarf hamster pups

In mammalian cross-species hybrids, parameters of voice calls, produced by vocal fold vibrations, are intermediate between parental species. Inheritance of ultrasonic calls, produced by whistle mechanism, is unstudied for hybrids. We examined 4000 pup ultrasonic isolation-induced calls for peak power of call fundamental frequency and for call duration in 4–8-day-old captive hamsters of four Study Groups: pure Phodopus sungorus; pure P. campbelli of two populations (Mongolian and Kosh-Agach) and hybrids between male P. sungorus and female P. campbelli (Kosh-Agach). All Study Groups produced two categories of ultrasonic calls: Low-Frequency centered around 41 kHz and High-Frequency centered around 60 kHz, but in different percentages. Between populations, only Low-Frequency calls were shorter and higher-frequency in Mongolian P. campbelli. Between species, only High-Frequency calls were shorter and higher-frequency in P. sungorus. In hybrids, Low-Frequency calls were shorter and lower-frequency than in either parental species, whereas High-Frequency calls were longer and lower-frequency in hybrids than in pure P. sungorus but similar with another parental species. We discuss that interspecific hybridization may give rise to offspring with new properties of ultrasonic calls.

Anthropogenic noise decreases activity and calling behavior in wild mice

Background Animals rely on sound to mediate a myriad of daily activities, and anthropogenic noise is a pollutant that alters the natural soundscape within which they are active. As human infrastructure expands, broadband anthropogenic noise increases, which can affect behaviors of free-living nocturnal animals. Mice are nocturnal animals that produce ultrasonic calls as part of their behavioral repertoire. Methods We assessed effects of anthropogenic and natural noise on the behaviors of wild deer mice (Peromyscus maniculatus) and woodland jumping mice (Napaeozapus insignis), two species of mice that produce ultrasonic calls. We measured activity, foraging behavior at a foraging tray, and calling behavior to broadcasts of natural and anthropogenic noise, compared to a baseline with no broadcasting, at 25 focal areas in the Southern Appalachian Mountain Range of North Carolina, USA. Results Deer mice exposed to anthropogenic noise spent less time in focal areas with broadcasted anthropogenic noise. Mice took longer to begin foraging in the presence of anthropogenic noise, they spent less time at the foraging tray, and left fewer husks but consumed the same number of seeds as mice exposed to natural noise. Deer mice were less likely than woodland jumping mice to be the first to enter the focal area and approach food when in the presence of anthropogenic noise. Both species produced few ultrasonic calls in the presence of broadcasted natural and anthropogenic noise compared to their baseline level of calling. We present the first calls recorded from woodland jumping mice. Conclusion Anthropogenic noise affects activity, foraging behavior, and calling behavior of nocturnal mice. Natural noise also affects the calling behavior of mice. Mouse species respond differently to anthropogenic noise, with deer mice appearing more sensitive to anthropogenic noise than woodland jumping mice. Responses to noise could have important effects on the ecology of mice and these two species respond differently. Species differences should be considered when mitigating the effects of noise in conservation ecology.

Batting around a new idea: attracting insect buffets for endangered bats

During the autumn months, many bat species face challenges to find sufficient food and generate the large fat stores they require to survive winter. However, North American bats face an additional challenge: a fungal disease called white-nose syndrome. The disease causes bats to arouse too frequently during hibernation, eventually starving to death before spring. Twelve bat species are currently affected, and some are listed as Endangered in either the USA or Canada. Winifred Frick, from Bat Conservation International and the University of California, Santa Cruz, USA, with a team of collaborators from various US and Canadian institutions, set out to test a new conservation approach that could protect bats by increasing the number of flying insects in an area, allowing the animals to gorge before and after hibernation.To test their approach, the team chose caves in the Upper Peninsula of Michigan, USA, which were known to have ∼100 remaining hibernating little brown bats (Myotis lucifugus) after declines from white-nose syndrome. They then monitored five caves in the autumn of 2019 (1 September–4 October), when the bats were fattening for hibernation, and four caves in the spring of 2021 (19 April–25 May), when bats that survived white-nose syndrome were emerging from hibernation. To attract flying insects, Frick and colleagues placed a single UV light 250 m from each cave entrance, 3 m above the ground. In the autumn, the team alternated nights when the UV lights were on, to assess changes in the number of insects they attracted. However, in the spring, to determine whether the bats could learn to respond to changes in the numbers of insects available to dine on, some of the caves were provided with a UV light on each night, while others had no UV lights on.The team also measured insect abundance every night during the autumn but only once a week during the spring, using a funnel and bucket to trap insects that were then brought back to the lab to be weighted and identified. To monitor bat foraging activity at the caves, the researchers used bat detectors placed at the sites to record the bats’ ultrasonic calls while flying from sunset to sunrise each night during the autumn and spring. Back in the lab, the recordings were reviewed to identify and count the number of ‘feeding buzzes’ – the echolocation calls that bats make when successfully catching insects – to determine the bats’ foraging success rates.The researchers found that when the UV lights were on during the autumn fattening period, the bats successfully captured and ate three times more insects than on nights when the UV lights were off. Additionally, the team collected a far greater mass of insects (16.7 times more) on the UV illuminated nights. Additionally, in the spring, the hunting bats were 8.5 times more successful in capturing prey, and the mass of insects collected by the team was 26.1 times greater at the locations where the UV lights were on than at the locations where the UV lights were off. These results show that UV lights in the autumn and spring increase the insect abundance for bats to eat, and that bats learn to forage more when there is increased food availability.Overall, Frick and colleagues have successively shown a novel approach for artificially attracting greater numbers of insects for bats to feast on during the critical spring and autumn periods. The results from the study could aid in the recovery of bat populations from white-nose syndrome and inspire wider conservation approaches for critically endangered hibernating bats in North America.

Cross-fostering effects on ultrasonic calls in two gerbil species

Cross-fostering effects on ultrasonic calls in two gerbil species

Mouse and rat ultrasonic vocalizations in neuroscience and neuropharmacology: State of the art and future applications.

Mice and rats emit ultrasonic vocalizations (USVs), which may express their arousal and emotional states, to communicate with each other. There is continued scientific effort to better understand the functions of USVs as a central element of the rodent behavioral repertoire. However, studying USVs is not only important because of their ethological relevance, but also because they are widely applied as a behavioral readout in various fields of biomedical research. In mice and rats, a large number of experimental models of brain disorders exist and studying the emission of USVs in these models can provide valuable information about the health status of the animals and the effectiveness of possible interventions, both environmental and pharmacological. This review (i) provides an updated overview of the contexts in which ultrasonic calling behaviour of mice and rats has particularly high translational value, and (ii) gives some examples of novel approaches and tools used for the analysis of USVs in mice and rats, combining qualitative and quantitative methods. The relevance of age and sex differences as well as the importance of longitudinal evaluations of calling and non-calling behaviour is also discussed. Finally, the importance of assessing the communicative impact of USVs in the receiver, that is, through playback studies, is highlighted.

Sex Differences in Behavior and Learning Abilities in Adult Rats

Laboratory rats have excellent learning abilities and are often used in cognitive neuroscience research. The majority of rat studies are conducted on males, whereas females are usually overlooked. Here, we examined sex differences in behavior and tactile sensitivity in littermates during adulthood (5.8-7.6 months of age). We used a battery of behavioral tests, including the 2% sucrose preference test (positive motivation), a free-choice paradigm (T-maze, neutral situation), and associative fear-avoidance learning (negative motivation, aversive situation). Tactile perception was examined using the von Frey test (aversive situation). In two aversive situations (von Frey test and avoidance learning), females were examined during the diestrus stage of the estrous cycle, and ultrasonic vocalization was recorded in both sexes. It was found that (1) females, but not males, lost their body weight on the first day of the sucrose preference test, suggesting sex differences in their reaction to environmental novelty or in metabolic homeostasis; (2) the tactile threshold in females was lower than in males, and females less frequently emitted aversive ultrasonic calls; (3) in the avoidance learning task, around 26% of males (but no females) were not able to learn and experienced frizzing. Overall, the performance of associative fear-avoidance in males was worse than in females. In general, females demonstrated higher abilities of associative learning and less persistently emitted aversive ultrasonic calls.

Playback of broadband vocalizations of female mice suppresses male ultrasonic calls.

Although male vocalizations during opposite- sex interaction have been heavily studied as sexually selected signals, the understanding of the roles of female vocal signals produced in this context is more limited. During intersexual interactions between mice, males produce a majority of ultrasonic vocalizations (USVs), while females produce a majority of human-audible squeaks, also called broadband vocalizations (BBVs). BBVs may be produced in conjunction with defensive aggression, making it difficult to assess whether males respond to BBVs themselves. To assess the direct effect of BBVs on male behavior, we used a split-cage paradigm in which high rates of male USVs were elicited by female presence on the other side of a barrier, but which precluded extensive male-female contact and the spontaneous production of BBVs. In this paradigm, playback of female BBVs decreased USV production, which recovered after the playback period. Trials in which female vocalizations were prevented by the use of female bedding alone or of anesthetized females as stimuli also showed a decrease in response to BBV playback. No non-vocal behaviors declined during playback, although digging behavior increased. Similar to BBVs, WNs also robustly suppressed USV production, albeit to a significantly larger extent. USVs suppression had two distinct temporal components. When grouped in 5-second bins, USVs interleaved with bursts of stimulus BBVs. USV suppression also adapted to BBV playback on the order of minutes. Adaptation occurred more rapidly in males that were housed individually as opposed to socially for a week prior to testing, suggesting that the adaptation trajectory is sensitive to social experience. These findings suggest the possibility that vocal interaction between male and female mice, with males suppressing USVs in response to BBVs, may influence the dynamics of communicative behavior.

Long-term behavioral effects of prenatal stress in the Fmr1-knock-out mouse model for fragile X syndrome.

Fragile X syndrome (FXS) is a major neurodevelopmental disorder and the most common monogenic cause of autism spectrum disorder (ASD). FXS is caused by a mutation in the X-linked FMR1 gene leading to the absence of the FMRP protein, inducing several behavioral deficits, including motor, emotional, cognitive, and social abnormalities. Beside its clear genetic origins, FXS can be modulated by environmental factors, e.g., stress exposure: indeed the behavioral phenotype of FXS, as well as of ASD patients can be exacerbated by the repeated experience of stressful events, especially early in life. Here we investigated the long-term effects of prenatal exposure to unpredictable chronic stress on the behavioral phenotype of the Fmr1-knock-out (KO) mouse model for FXS and ASD. Mice were tested for FXS- and ASD-relevant behaviors first at adulthood (3 months) and then at aging (18 months), in order to assess the persistence and the potential time-related progression of the stress effects. Stress induced the selective emergence of behavioral deficits in Fmr1-KO mice that were evident in spatial memory only at aging. Stress also exerted several age-specific behavioral effects in mice of both genotypes: at adulthood it enhanced anxiety levels and reduced social interaction, while at aging it enhanced locomotor activity and reduced the complexity of ultrasonic calls. Our findings underline the relevance of gene-environment interactions in mouse models of neurodevelopmental syndromes and highlight the long-term behavioral impact of prenatal stress in laboratory mice.

Neuroethology of sound localization in anurans.

Albert Feng pioneered the study of neuroethology of sound localization in anurans by combining behavioral experiments on phonotaxis with detailed investigations of neural processing of sound direction from the periphery to the central nervous system. The main advantage of these studies is that many species of female frogs readily perform phonotaxis towards loudspeakers emitting the species-specific advertisement call. Behavioral studies using synthetic calls can identify which parameters are important for phonotaxis and also quantify localization accuracy. Feng was the first to investigate binaural processing using single-unit recordings in the first two auditory nuclei in the central auditory pathway and later investigated the directional properties of auditory nerve fibers with free-field stimulation. These studies showed not only that the frog ear is inherently directional by virtue of acoustical coupling or crosstalk between the two eardrums, but also confirmed that there are extratympanic pathways that affect directionality in the low-frequency region of the frog's hearing range. Feng's recordings in the midbrain also showed that directional information is enhanced by cross-midline inhibition. An important contribution toward the end of his career involved his participation in neuroethological research with a team of scientists working with frogs that produce ultrasonic calls.

Norway rats help social partners in need in response to ultrasonic begging signals

AbstractBegging signals are widespread in animals and have been extensively studied in the context of brood care. In contrast, the functionality of begging signals expressed among adults has not yet been scrutinised experimentally. Norway rats (Rattus norvegicus) have been shown to express 50‐kHz calls towards a social partner when in need of help to obtain food. Here, we tested with help of playbacks whether focal subjects exposed to a high rate of ultrasonic calls would increase their helping rate towards their social partner. The propensity to help a partner was significantly enhanced when the focal rats were exposed to playback with a doubled number of calls relative to baseline conditions. To the best of our knowledge, this provides the first evidence of increased help in response to an experimentally altered begging signal in adult animals.

Quantification of Behavioral Deficits in Developing Mice With Dystonic Behaviors.

Converging evidence from structural imaging studies in patients, the function of dystonia-causing genes, and the comorbidity of neuronal and behavioral defects all suggest that pediatric-onset dystonia is a neurodevelopmental disorder. However, to fully appreciate the contribution of altered development to dystonia, a mechanistic understanding of how networks become dysfunctional is required for early-onset dystonia. One current hurdle is that many dystonia animal models are ideally suited for studying adult phenotypes, as the neurodevelopmental features can be subtle or are complicated by broad developmental deficits. Furthermore, most assays that are used to measure dystonia are not suited for developing postnatal mice. Here, we characterize the early-onset dystonia in Ptf1a Cre ;Vglut2 fl/fl mice, which is caused by the absence of neurotransmission from inferior olive neurons onto cerebellar Purkinje cells. We investigate motor control with two paradigms that examine how altered neural function impacts key neurodevelopmental milestones seen in postnatal pups (postnatal day 7-11). We find that Ptf1a Cre ;Vglut2 fl/fl mice have poor performance on the negative geotaxis assay and the surface righting reflex. Interestingly, we also find that Ptf1a Cre ;Vglut2 fl/fl mice make fewer ultrasonic calls when socially isolated from their nests. Ultrasonic calls are often impaired in rodent models of autism spectrum disorders, a condition that can be comorbid with dystonia. Together, we show that these assays can serve as useful quantitative tools for investigating how neural dysfunction during development influences neonatal behaviors in a dystonia mouse model. Our data implicate a shared cerebellar circuit mechanism underlying dystonia-related motor signs and social impairments in mice.

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

Ultrasonic vocalizations (USVs) are a major tool for assessing social communication in laboratory mice during their entire lifespan. At adulthood, male mice preferentially emit USVs toward a female conspecific, while females mostly produce ultrasonic calls when facing an adult intruder of the same sex. Recent studies have developed several sophisticated tools to analyze adult mouse USVs, especially in males, because of the increasing relevance of adult communication for behavioral phenotyping of mouse models of autism spectrum disorder (ASD). Little attention has been instead devoted to adult female USVs and impact of sex differences on the quantitative and qualitative characteristics of mouse USVs. Most of the studies have also focused on a single testing session, often without concomitant assessment of other social behaviors (e.g., sniffing), so little is still known about the link between USVs and other aspects of social interaction and their stability/variations across multiple encounters. Here, we evaluated the USVs emitted by adult male and female mice during 3 repeated encounters with an unfamiliar female, with equal or different pre-testing isolation periods between sexes. We demonstrated clear sex differences in several USVs' characteristics and other social behaviors, and these were mostly stable across the encounters and independent of pre-testing isolation. The estrous cycle of the tested females exerted quantitative effects on their vocal and non-vocal behaviors, although it did not affect the qualitative composition of ultrasonic calls. Our findings obtained in B6 mice, i.e., the strain most widely used for engineering of transgenic mouse lines, contribute to provide new guidelines for assessing ultrasonic communication in male and female adult mice.

Alterations between high and low-avoidance lines of Hatano rats in learning behaviors, ultrasonic vocalizations, and histological characteristics in hippocampus and amygdala

Growing evidence supports interactions between anxiety and cognitive function. The primary object of this study was to elucidate whether high-avoidance (HAA) and low-avoidance (LAA) strains of Hatano rats are suitable for the analysis of interactions between the formation of long-term memory and emotional reactivity. The learning/memory ability of Hatano rats and their Sprague–Dawley (SD) ancestors was evaluated using contextual fear conditioning, Y-maze, and Barnes maze tests from 8 weeks of age. Ultrasonic vocalizations were recorded and analyzed during contextual fear conditioning. In a separate experiment, rat brains were sampled 90 min after the first context test and subjected to Nissl staining and c-fos immunostaining. The duration of freezing and number of 22 kHz ultrasonic vocalizations were decreased in LAA compared with HAA and SD rats during the first and second context tests of contextual fear conditioning. The HAA rats did not show preferences for quadrants during the Barnes maze probe test, whereas the SD and LAA rats spent significantly more time in the quadrant where the goals had been placed. There was no difference among the strains in short-term spatial memory as shown by the Y-maze test. Decreases were found in the number of c-fos+ cells as well as the volume of some hippocampal regions in the HAA rats compared to SD and LAA rats. By contrast, the volume of the basolateral amygdala was bigger in the HAA than the other strains. On the basis of the 22 kHz ultrasonic calls and literature regarding Syracuse rats, the possibility that emotional reactivity influences contextual memory in Hatano strains was discussed. This emotional difference may be derived from structural and/or functional divergence in the hippocampus and amygdala between the strains. The cause of strain-related differences in long-term spatial learning was difficult to elucidate because there are several possible explanations, including differences in memory and/or the interference of hyperactivity during the Barnes maze test.

Rat 50 kHz Trill Calls Are Tied to the Expectation of Social Interaction.

Rats emit a variety of calls in the 40–80 kHz range (50 kHz calls). While these calls are generally associated with positive affect, it is unclear whether certain calls might be used selectively in certain contexts. To examine this, we looked at ultrasonic calls in 30–40 day old male rats during the expectation of either play or food, both of which are reinforcing. Behavior and vocalizations were recorded while rats were in a test chamber awaiting the arrival of a play partner or food over seven days of testing. Control groups were included for the non-specific effects of food deprivation and social isolation. Play reward led to an increase in 50 kHz vocalizations, generally, with specific increases in trill and “trill with jump” calls not seen in other groups. Expectation of food reward did not lead to a significant increase in vocalizations of any type, perhaps due to the young age of our study group. Further, rats that were food deprived for the food expectation study showed markedly lower calls overall and had a different profile of call types compared to rats that were socially isolated. Taken together, the results suggest that trill-associated calls may be used selectively when rats are socially isolated and/or expecting a social encounter.