Testing social acoustic memory in rats: Effects of stimulus configuration and long-term memory on the induction of social approach behavior by appetitive 50-kHz ultrasonic vocalizations

Rats emit distinct types of ultrasonic vocalizations (USVs), which serve as situation-dependent affective signals. In appetitive situations, such as rough-and-tumble-play, high-frequency 50-kHz USVs occur, whereas low-frequency 22-kHz USVs can be observed in aversive situations, such as social defeat. USVs serve distinct communicative functions and induce call-specific behavioral responses in the receiver. While aversive 22-kHz USVs serve as alarm calls and induce behavioral inhibition, appetitive 50-kHz USVs have a pro-social communicative function and elicit social approach behavior, supporting the notion that they serve as social contact calls to (re)establish or maintain contact among conspecifics. The aim of the present study was to use the rat's ability to communicate in the ultrasonic range via high-frequency 50-kHz USVs in order to develop a test for social acoustic memory in rats with relevance for human verbal memory. Verbal learning and memory is among the seven cognitive domains identified as commonly deficient in human schizophrenia patients, but particularly difficult to model. We therefore tested whether the induction of social approach behavior by playback of appetitive 50-kHz USVs is dependent on (1) acoustic stimulus configuration and (2) social long-term memory, and whether (3) social long-term memory effects can be blocked by the administration of scopolamine, a muscarinic acetylcholine antagonist producing amnesia. Results show that social approach behavior in response to playback of natural 50-kHz USVs depends on acoustic stimulus configuration and occurs only when sound energy is concentrated to a critical frequency band in the ultrasonic range. Social approach behavior was detected during the first exposure to playback of 50-kHz USVs, whereas no such response was observed during the second exposure 1week later, indicating a stable memory trace. In contrast, when memory formation was blocked by i.p. administration of scopolamine (0.5mg/kg or 1.5mg/kg) immediately after the first exposure, rats displayed social approach behavior during the second exposure as well. Induction of social approach behavior in response to repeated playback of natural 50-kHz USVs may therefore provide a new and rather unique approach for testing social acoustic memory in rats with relevance to human verbal memory.     

Ultrasonic vocalizing by adult female mice (Mus musculus)

Though an extensive body of literature exists concerning the emission of 70-kHz ultrasonic vocalizations by adult male mice (Mus musculus), almost nothing is known about the ultrasonic vocalizing of adult females. A series of five experiments was conducted to examine adult female mouse ultrasound emission. Results indicated female ultrasonic vocalizing to be typically displayed among female mouse dyads and comparable to the ultrasonic vocalizing levels obtained among male-female pairs. Genotypically based variations in the production of ultrasounds by females were also noted. Like male mice, socially naive females readily ultrasonically vocalized to anesthetized female conspecifics and rarely vocalized to anesthetized males. Unlike males, socially experienced females emitted few ultrasounds to either female urine or female-soiled cage shavings. Although social experience served to increase the ultrasound emission of male mice to female sex cues, the production of ultrasounds by females to these cues was decreased by social experience. Implications with regard to the existence of a behavioral/functional sexual dimorphism in adult mouse ultrasonic vocalizing are discussed.     

Relation between 22-kHz ultrasonic signals and sociosexual behavior in rats

The copulatory performance of male rats, tested in a large seminaturalistic environment, was assessed to determine the relation between 22-kHz ultrasonic vocalizations and a range of sociosexual behaviors. The male rats were tested until sexual exhaustion. Such ultrasonic signals were shown to occur in a wider range of sociosexual circumstances than previously reported; for example, the calls occcurred in particular social circumstances during the preejaculatory period as well as during the postejaculatory interval. There was no consistent evidence that the emission of this call during the postejaculatory period consistently functions to keep the female away from the male. The nature and occurrence of postejaculatory ultrasonic signals showed increasing variability in successive ejaculatory series. The results of this and previous studies are interpreted within a semiotic theory of communication. The 22-kHz call is described as a message that makes available the information that the sender is in a socially depressed and withdrawn state.     

The persistence of some effects of handling in infancy on the behaviour of the adult rat

An attempt was made to discover how persistent the effects of handling before weaning are on activity, grooming, emotional elimination and learning in adult rats; whether there is an interaction of the effects of handling and the effects of cage size on the behaviour of rats; and whether the daily removal of the mother during nursing has any effects on the pups.

Since all differences between handled and undisturbed animals found at 50 or 100 days were found at 220 days, it was concluded that the effects of handling in infancy are persistent. The hypothesis that cage size and removal of the mother during nursing significantly affect adult behaviour was not supported by the results.     

Juvenile stress potentiates aversive 22-kHz ultrasonic vocalizations and freezing during auditory fear conditioning in adult male rats

Traumatic experiences that occur during adolescence can render individuals vulnerable to mood and anxiety disorders. A model in juvenile rats (age: 27-29 days) was developed previously to study the long-term effects of adolescent stress exposure on behaviour and physiology. This paradigm, termed juvenile stress, involves subjecting juvenile rats to different stressors on consecutive days over a 3-day period. Here, we investigated the effects of the juvenile stress paradigm on freezing behaviour and aversive 22-kHz ultrasonic vocalizations (USVs) during auditory fear conditioning in adult male rats (age: 68-90 days). We found that rats previously subjected to juvenile stress increased aversive 22-kHz USVs (total calls and time spent calling) compared with controls during fear-conditioning training. The acoustic USV parameters between control and juvenile stress rats were largely equivalent, including duration, peak frequency and amplitude. While rats did not differ in freezing behaviour during fear conditioning, juvenile stress rats exhibited greater cue-conditioned freezing upon testing 24 h later. Our results show that juvenile stress elicited different long-term changes in freezing and aversive USVs during fear conditioning. Furthermore, they highlight the importance of assessing USVs to detect experience-dependent differences between control and stress-exposed animals which are not detectable by measuring visible behaviour.     

Ultrasonic vocalizations as indices of affective states in rats

Adult rats spontaneously vocalize in ultrasonic frequencies. Although these ultrasonic vocalizations (USVs) have been described as by-products of locomotor activity or social signals, accumulating evidence suggests that they may also index anticipatory affective states. Converging ethological, pharmacological, and brain stimulation research indicates that whereas long low-frequency (> 0.3-s, approximately 22-kHz) USVs occur during anticipation of punishment or avoidance behavior, short, high-frequency (< 0.3-s, approximately 50-kHz) USVs typically occur during anticipation of reward or approach behavior. Thus, long 22-kHz USVs may index a state of negative activation, whereas short, 50-kHz USVs may instead index a state of positive activation. This hypothesis has theoretical implications for understanding the brain circuitry underlying mammalian affective states and clinical applicability for modeling hedonic properties of different psychotropic compounds.     

Analysis of ultrasonic vocalizations emitted by residents during aggressive encounters among rats (Rattus norvegicus)

This investigation was concerned with the extent to which aggressive resident rats emit 40-70-kHz vocalizations and the effect of these signals on intruders. In Experiment 1, deafened and intact intruder males were given two encounters with resident animals. Deafened intruders engaged in a higher duration of immobile or freezing postures than intact animals. Experiment 2 indicated that the augmentation of freezing found among deafened intruders was not due to an inability to detect ultrasounds made by residents since intruders encountering devocalized resident males showed no reliable differences in specific motor patterns from intruders paired with intact residents. The results further demonstrated that 40-70-kHz vocalizations are produced almost entirely by intruding animals since there were no significant changes in occurrence of these calls when resident males were devocalized. Under the constraints of the testing procedures employed, the role of ultrasonic communication during the initial formation of agonistic relations could not be determined experimentally.     

Analysis of ultrasonic vocalizations emitted by intruders during aggressive encounters among rats (Rattus norvegicus)

This investigation was concerned with the identification of the ultrasonic vocalizations produced by intruders during aggressive interactions and the role of these signals in agonistic behavior of rats. In the first experiment, experienced resident males were paired with both devocalized and intact vocalizing naive intruder males. Devocalization of the intruder males resulted in a drastic decrease in 50-kHz vocalizations and the elimination of all 22-kHz vocalizations. This almost total absence of ultrasonic vocalizations was not accompanied by any change in resident aggressive behavior or intruder defensive and submissive behavior. In a second experiment, naive intruders were tested with either deafened or intact resident males. Similarly, preventing residents from hearing intruder ultrasounds had no detectable effect on any aggressive behavior. These experiments are not consistent with the correlative evidence that intruder-produced 22-kHz vocalizations inhibit the aggressive behavior of the resident. The results also show that most of the ultrasonic vocalizations emitted during aggressive encounters are probably produced by the intruder.     

Playful defensive responses in adult male rats depend on the status of the unfamiliar opponent

Adult male rats reared as pairmates from weaning were tested in a neutral arena with both members of another pair (one at a time). The unfamiliar pairs were found to engage in play fighting, although they were more likely to escalate the encounter into serious fighting than were pairs of familiar rats. Based on their within‐home pair behavior, each pairmate was designated as a dominant or a subordinate. When the test encounters between unfamiliar males were analyzed with regard to whether the pairings consisted of two dominants, two subordinates, or a mixed pair, the pattern of play fighting was found to be attenuated. Both dominants and subordinates were more likely to initiate playful encounters, to respond defensively during these encounters, and to do so using adult‐typical tactics of defense when paired with an unfamiliar rat that was dominant in its home cage. The mechanisms by which the home status of unfamiliar male rats can be identified by another male are discussed, particularly with regard to the role that play fighting may serve for this function. It is concluded that the data support the hypothesis that play fighting can be used by adult rats for social testing, which in this case seems to involve ascertaining the opponent's fighting capability. Aggr. Behav. 25:141–152, 1999. © 1999 Wiley‐Liss, Inc.     

Aggression in domesticated rats reared in a burrow-digging environment

Male and female rats were raised individually in either large boxes filled with soil (burrow environment) or laboratory cages of comparable size (cage environment). In a test of intruder-elicited aggression, intruders exposed to burrow-environment residents lost more body weight, were more likely to have empty stomachs, and received more wounds during a 24 hour aggression test. The results indicate that the physical environment during the period from weaning to maturity is an important determinant of aggression intensity in adult rats.     

Ultrasonic vocalizations by male house mice (Mus musculus) to novel odors: roles of infant and adult experience

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)     

On the function of redfronted lemur’s close calls

In order to maintain group cohesion, many social mammals and birds regularly produce close calls. In some primate species, close calls appear to have a dual function: calls addressed at a broad class of targets serve to maintain group cohesion, whereas the same calls directed at a specific target serve to regulate subsequent social interactions. Redfronted lemurs (Eulemur rufifrons) produce different types of close calls: grunts, long grunts, hoos and meows. In order to study the function of these calls, we conducted focal observations and vocal recordings from eight adult males and females out of four social groups in Kirindy Forest, Western Madagascar. Redfronted lemurs produce long grunts, hoos and meows at relatively low rates during foraging, resting or group movements, respectively. Grunts were given most often and more or less constantly during foraging and traveling. Calling rate increased when the risk of separation increased and may thus promote group cohesion. Grunts given during approaches of other group members resulted more often in friendly interactions than approaches that were not accompanied by a grunt. Thus, redfronted lemurs produce specific but also generic contact calls, whereas the latter calls have a dual function that varies depending on the addressed audience: they act as an auditory beacon to maintain group cohesion and serve as signals of benign intent to avoid costly conflicts and facilitating social interactions.     

Ultrasonic vocalizations during sex behavior of male house mice (Mus musculus): a description

After placing a female house mouse into the home cage of a male, the occurrences of four behaviors were recorded on separate channels of an event recorder: (1) male sniffing female, (2) male mounting female, (3) male intromitting female, and (4) 70-kHz vocalizations. The amount of vocalizing was greatest shortly after pairing and was associated with the male sniffing the female. After the male began mounting, vocalizations also were associated with mounting. Vocalizations were recorded during intromissions and occasionally occurred coincident with pelvic thrusts. Very few vocalizations were detected when the male was not sniffing or mounting the female. Vocalizations ceased following ejaculation but typically resumed several minutes before the resumption of another mounting sequence. Thus 70-kHz vocalizations appear to be closely linked to male sexual arousal.     

Social transmission of Pavlovian fear: fear-conditioning by-proxy in related female rats

Pairing a previously neutral conditioned stimulus (CS; e.g., a tone) to an aversive unconditioned stimulus (US; e.g., a foot-shock) leads to associative learning such that the tone alone will elicit a conditioned response (e.g., freezing). Individuals can also acquire fear from a social context, such as through observing the fear expression of a conspecific. In the current study, we examined the influence of kinship/familiarity on social transmission of fear in female rats. Rats were housed in triads with either sisters or non-related females. One rat from each cage was fear conditioned to a tone CS+ shock US. On day two, the conditioned rat was returned to the chamber accompanied by one of her cage mates. Both rats were allowed to behave freely, while the tone was played in the absence of the foot-shock. The previously untrained rat is referred to as the fear-conditioned by-proxy (FCbP) animal, as she would freeze based on observations of her cage-mate’s response rather than due to direct personal experience with the foot-shock. The third rat served as a cage-mate control. The third day, long-term memory tests to the CS were performed. Consistent with our previous application of this paradigm in male rats (Bruchey et al. in Behav Brain Res 214(1):80–84, 2010), our results revealed that social interactions between the fear conditioned and FCbP rats on day two contribute to freezing displayed by the FCbP rats on day three. In this experiment, prosocial behavior occurring at the termination of the cue on day two was significantly greater between sisters than their non-sister counterparts, and this behavior resulted in increased freezing on day three. Our results suggest that familiarity and/or kinship influences the social transmission of fear in female rats.     

Enduring deficits in contextual and auditory fear conditioning after adolescent, not adult, social instability stress in male rats

Adolescence is a time of developmental changes and reorganization in the brain and stress systems, thus, adolescents may be more vulnerable than adults to the effects of chronic mild stressors. Most studies, however, have not directly compared stress experienced in adolescence to the same stress experience in adulthood. In the present study, adolescent (n=46) and adult (n=48) male rats underwent 16 days of social instability stress (daily 1h isolation and change of cage partners) or were non-stress controls. Rats were then tested on the strength of acquired contextual and cued fear conditioning, as well as extinction learning, beginning either the day after the stress procedure or 3 weeks later. No difference was found among the groups during the Training Phase of conditioning. Irrespective of the time between the social stress experience and fear conditioning, rats stressed in adolescence had decreased context and cue memory, and cue generalization compared to control rats, as measured by the percentage of time spent freezing in tests. Social instability stress in adulthood had no effect on any measure of fear conditioning. The results support the hypothesis that adolescence is a time of heightened vulnerability to stressors.     

Ultrasounds produced by rats accompany decreases in intraspecific fighting

Male intruder rats were placed individually into the cage of an established resident on 2 occasions separated by a 7–8 day interval. Residents readily attacked intruders and both animals lost weight during the first encounter. In contrast, no serious fighting occurred on the second encounter, and both intruders and residents maintained their body weight during the 24-hr test. Observation of the intruder's behavior during the first 30 min of each encounter indicated that defensive-submissive postures represent a response to an attack that only temporarily inhibits aggression whereas the emission of 22 kHz calls by the intruder is associated with a relatively permanent decrease in the resident animal's aggressive response.     

Interfemale aggression in adult bank voles (Clethrionomys glareolus)

Agonistic interaction may play a large role in shaping the social structure and spatial distribution of rodents such as Clethrionomys glareolus. Aggressive and non-aggressive behaviors of adult bank vole females were investigated with a home cage test during 10 min sessions. The number of attacks was significantly higher in virgin females kept with intact adult males than in singly reared females or those exposed to male urine. Bulbectomy of the tested females completely inhibited the male stimulating effect. This indicates that males influence the behavior of females by activating their olfactory systems. Aggressive behavior in pregnant or lactating females, which is linked to maternal behavior, was also tested. Female bank voles between 11 and 17 days of pregnancy or lactating at 3–4 days or 9–10 days attacked their partners significantly more frequently than virgins did. Females that had direct contact with males, even if they were not fertilized, appeared to be more successful in same-sex competition. Aggr. Behav. 24:53–61, 1998. © 1998 Wiley-Liss, Inc.     

Social enrichment improves social recognition memory in male rats

The social environment is thought to have a strong impact on cognitive functions. In the present study, we investigated whether social enrichment could affect rats’ memory ability using the “Different Objects Task (DOT),” in which the levels of memory load could be modulated by changing the number of objects to be remembered. In addition, we applied the DOT to a social discrimination task using unfamiliar conspecific juveniles instead of objects. Animals were housed in one of the three different housing conditions after weaning [postnatal day (PND) 21]: social-separated (1 per cage), standard (3 per cage), or social-enriched (10 per cage) conditions. The object and social recognition tasks were conducted on PND 60. In the sample phase, the rats were allowed to explore a field in which 3, 4, or 5 different, unfamiliar stimuli (conspecific juveniles through a mesh or objects) were presented. In the test phase conducted after a 5-min delay, social-separated rats were able to discriminate the novel conspecific from the familiar ones only under the condition in which three different conspecifics were presented; social-enriched rats managed to recognize the novel conspecific even under the condition of five different conspecifics. On the other hand, in the object recognition task, both social-separated and social-enriched rats were able to discriminate the novel object from the familiar ones under the condition of five different objects. These results suggest that social enrichment can enhance social, but not object, memory span.     

Overt behavior and ultrasonic vocalization in a fear conditioning paradigm: a dose-response study in the rat

The behavioral analysis of laboratory rats is usually confined to the level of overt behavior, like locomotion, behavioral inhibition, instrumental responses, and others. Apart from such visible outcome, however, behaviorally relevant information can also be obtained when analyzing the animals' ultrasonic vocalization, which is typically emitted in highly motivational situations, like 22-kHz calls in response to acute or conditioned threat. To further investigate such vocalizations and their relationship with overt behavior, we tested male Wistar rats in a paradigm of Pavlovian fear conditioning, where a tone stimulus (CS) was preceding an aversive foot-shock (US) in a distinct environment. Importantly, the shock dose was varied between groups (0-1.1 mA), and its acute and conditioned outcome were determined. The analysis of visible behavior confirms the usefulness of immobility as a measure of fear conditioning, especially when higher shock doses were used. Rearing and grooming, on the other hand, were more useful to detect conditioned effects with lower shock levels. Ultrasonic vocalization occurred less consistently than changes in overt behavior; however, dose-response relationships were also observed during the phase of conditioning, for example, in latency, call rate and lengths, intervals between calls, and sound amplitude. Furthermore, total calling time (and rate) were highly correlated with overt behavior, namely behavioral inhibition as measured through immobility. These correlations were observed during the phase of fear conditioning, and the subsequent tests. Importantly, conditioned effects in overt behavior were observed, both, to the context and to the CS presented in this context, whereas conditioned vocalization to the context was not observed (except for one rat). In support and extent of previous results, the present data show that a detailed analysis of ultrasonic vocalization can substantially broaden and refine the spectrum of analysis in behavioral work with rats, since it can provide information about situational-, state-, and subject-dependent factors which are partly distinct from what is visible to the experimenter.     

Intermittent exposure to social defeat and open-field test in rats: acute and long-term effects on ECG,body temperature and physical activity

This study investigated the effects of exposure to an intermittent homotypic stressor on: (i) habituation of acute autonomic responsivity (i.e. cardiac sympathovagal balance and susceptibility to arrhythmias), and (ii) circadian rhythmicity of heart rate, body temperature, and physical activity. After implantation of a transmitter for the radiotelemetric recording of electrocardiogram (ECG), body temperature and physical activity, adult male rats (Rattus norvegicus, Wild Type Groningen strain) were repeatedly exposed (10 consecutive times, on alternate days) to either a social stressor (defeat by a con-specific, n = 15) or an open-field, control challenge (transfer to a new cage; n = 8). ECGs, body temperature and physical activity were continuously recorded in baseline, test and recovery periods (each lasting 15 min), at the 1st and 10th episodes of both defeat and open-field challenge. The circadian rhythms of heart rate, body temperature and physical activity were monitored before (5 days), during (16 days) and after (21 days) the intermittent stress protocol. This study indicates that there is no clear habituation of either acute cardiac autonomic responsivity (as estimated by means of time-domain indexes of heart rate variability) or arrhythmia occurrence to a brief, intermittent, homotypic challenge, regardless of the nature of the stressor (social or non-social). On the other hand, rats exposed to social challenge also failed to show adaptation of acute temperature and activity stress responsiveness, whereas rats facing open-field challenge developed habituation of activity and sensitization of temperature responses. Repeated social challenge produced remarkable reductions of the heart rate circadian rhythm amplitude (this effect being significantly greater than that produced by intermittent open-field), but only minor changes in the daily rhythms of body temperature and physical activity.