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.

Fear Extinction and Predictive Trait-Like Inter-Individual Differences in Rats Lacking the Serotonin Transporter.

Anxiety disorders are associated with a failure to sufficiently extinguish fear memories. The serotonergic system (5-hydroxytryptamine, 5-HT) with the 5-HT transporter (5-HTT, SERT) is strongly implicated in the regulation of anxiety and fear. In the present study, we examined the effects of SERT deficiency on fear extinction in a differential fear conditioning paradigm in male and female rats. Fear-related behavior displayed during acquisition, extinction, and recovery, was measured through quantification of immobility and alarm 22-kHz ultrasonic vocalizations (USV). Trait-like inter-individual differences in novelty-seeking, anxiety-related behavior, habituation learning, cognitive performance, and pain sensitivity were examined for their predictive value in forecasting fear extinction. Our results show that SERT deficiency strongly affected the emission of 22-kHz USV during differential fear conditioning. During acquisition, extinction, and recovery, SERT deficiency consistently led to a reduction in 22-kHz USV emission. While SERT deficiency did not affect immobility during acquisition, genotype differences started to emerge during extinction, and during recovery rats lacking SERT showed higher levels of immobility than wildtype littermate controls. Recovery was reflected in increased levels of immobility but not 22-kHz USV emission. Prominent sex differences were evident. Among several measures for trait-like inter-individual differences, anxiety-related behavior had the best predictive quality.

Reduced emission of alarm 22-kHz ultrasonic vocalizations during fear conditioning in rats lacking the serotonin transporter.

Rats display a rich social behavioral repertoire. An important component of this repertoire is the emission of whistle-like calls in the ultrasonic range, so-called ultrasonic vocalizations (USV). Long low-frequency 22-kHz USV occur in aversive situations, including aggressive interactions, predator exposure, and electric shocks during fear conditioning. They are believed to reflect a negative affective state akin to anxiety and fear. A prominent theory suggests that 22-kHz USV function as alarm calls to warn conspecifics. Serotonin (5-hydroxytryptamine, 5-HT) is strongly implicated in the regulation of affective states, particularly anxiety and fear. A key component of the system is the 5-HT transporter (5-HTT, also known as SERT), regulating 5-HT availability in the synaptic cleft. In the present experiment, we studied the effects of SERT deficiency on overt fear-related behavior and alarm 22-kHz USV during fear conditioning in male and female rats. While overt fear-related behavior was not affected by SERT deficiency and sex, the emission of alarm 22-kHz USV was clearly reduced in homozygous SERT-/- but not heterozygous SERT+/- mutants, as compared to their wildtype SERT+/+ littermate controls. Genotype effects were particularly prominent in females. Females in general emitted fewer alarm 22-kHz USV than males. This supports the view that 22-kHz USV are, at least partly, independently regulated from anxiety or fear and as socially mediated alarm calls do not simply express a negative affective state. Reduced 22-kHz USV emission in rats lacking SERT might be due to social deficits in the use of 22-kHz USV as a socio-affective signal to warn conspecifics about threats.

Sex-dependent effects of Cacna1c haploinsufficiency on behavioral inhibition evoked by conspecific alarm signals in rats.

Deficits in processing social signals leads to reduced social functioning and is typically associated with neuropsychiatric disorders, including autism spectrum disorder, schizophrenia, and major depressive disorder. The cross-disorder risk gene CACNA1C is implicated in the etiology of all of these disorders and single-nucleotide polymorphisms within CACNA1C are ranked among the best replicated and most robust genetic findings from genome-wide association studies in psychiatry. Rats are highly social, live in large social groups, and communicate through ultrasonic vocalizations (USV), with low-frequency 22-kHz USV emitted in dangerous and often life-threating situations, such as predator exposure, serving an alarming function. In the present study, we applied an alarm 22-kHz USV playback paradigm to investigate the role of Cacna1c in socio-affective information processing in rats. Specifically, we assessed behavioral inhibition evoked by 22-kHz USV in constitutive heterozygous Cacna1c+/- females and males, as compared to wildtype Cacna1c+/+ littermate controls. To probe specificity, two sets of alarm 22-kHz USV were presented, i.e. 22-kHz USV elicited by predator urine exposure and 22-kHz USV emitted during a retention test on learned fear, together with acoustic control stimuli. Our results show that behavioral inhibition evoked by playback of alarm 22-kHz USV is robust and occurs in response to both sets, yet is modulated by Cacna1c in a sex-dependent manner. In male but not female rats, Cacna1c haploinsufficiency led to less pronounced and less specific behavioral inhibition, supporting the idea that Cacna1c haploinsufficiency results in a lower motivation and/or diminished capability to display appropriate responses to important socio-affective communication signals.

Predator odour but not TMT induces 22-kHz ultrasonic vocalizations in rats that lead to defensive behaviours in conspecifics upon replay.

Predator odours induce defensive behaviour in prey animals such as rats. The present study investigated (1) whether laboratory rats exposed to predator odours emit 22-kHz calls which may have an alarming function and (2) whether playback of such calls induces behavioural changes in conspecifics. For this, Sprague-Dawley rats were exposed to samples of fox and lion urine, as well as to the synthetic predator odour TMT. Despite that all odours induced defensive behaviour, only predator urine samples but not TMT were able to induce 22-kHz calls in a few rats. In a second experiment, naive rats were exposed to playback presentations of the 22-kHz calls recorded in the first experiment, as well as to phase-scrambled and frequency-shifted control stimuli. Low intensity playback presentations led to a reduction of locomotor activity during the presentation of the 22-kHz calls but not of the control stimuli. This effect was less specific under high intensity conditions. Taken together the present findings show that natural predator odours are able to induce emission of 22-kHz calls in rats and support the hypothesis that these calls have an alarming function.

Effects of anxiogenic drugs on the emission of 22- and 50-kHz ultrasonic vocalizations in adult rats.

RATIONALE: Adult rat 22-kHz vocalizations are often associated with alarm or distress, whereas a subset of 50-kHz calls is preferentially emitted in response to amphetamine and other rewarding stimuli. Whether any 50-kHz calls reflect anxiety is unknown. OBJECTIVE: To determine the effects of anxiogenic drugs on 50-kHz call rate and call subtype profile, in comparison with D-amphetamine. METHODS: Adult male rats received systemic amphetamine (1 mg/kg) three times several days before testing. Ultrasonic vocalizations were then recorded after acute intraperitoneal injection of amphetamine or one of five anxiogenic drugs: yohimbine (2.5 mg/kg), N-methyl-ß-carboline-3-carboxamide (FG 7142, 5 mg/kg), pentylenetetrazol (PTZ, 20 mg/kg), m-chlorophenylpiperazine (mCPP, 1 mg/kg), caffeine (25 mg/kg), or vehicle. RESULTS: The duration of immobility was increased by FG 7142, PTZ, and mCPP; this measure was unchanged by yohimbine and reduced by the locomotor stimulant drugs amphetamine and caffeine. Conversely, the 50-kHz call rate was reduced by FG 7142, PTZ and mCPP, and increased by caffeine and amphetamine. Overall, the most common 50-kHz call subtypes were flat, trill, step-up, and complex. Consistent with previous reports, amphetamine increased the relative prevalence of trill calls while reducing the relative prevalence of flat calls. Yohimbine and caffeine reduced flat call prevalence, whereas mCPP reduced trill call prevalence. No other shifts in the call profile were observed, and no anxiogenic drug induced 22-kHz calls. CONCLUSION: Anxiogenic drugs, as a class, did not uniformly alter the 50-kHz call rate or subtype profile. Amphetamine-induced effects on 50-kHz call rate and profile do not reflect anxiety.

Rodent ultrasonic communication: Male prosocial 50-kHz ultrasonic vocalizations elicit social approach behavior in female rats (Rattus norvegicus).

Rats emit distinct types of ultrasonic vocalizations (USV), which serve as situation-dependent affective signals with important communicative functions. Low-frequency 22-kHz USV typically occur in aversive situations, such as social defeat, whereas high-frequency 50-kHz USV can be observed in appetitive situations, like rough-and-tumble-play in juveniles or mating in adults. The 2 main USV types serve distinct communicative functions and induce call-specific behavioral responses in the receiver. While 22-kHz USV probably serve as alarm calls, 50-kHz USV appear to serve a prosocial communicative function in the sexual and the nonsexual context. In the sexual context, however, this view has recently been challenged by playback studies where only very limited behavioral changes were observed in response to prosocial 50-kHz USV. The aim of the present study was therefore to test whether female rats display social approach behavior in response to male prosocial 50-kHz USV by means of our established playback paradigm. To this aim, we exposed female rats to playback of the following 2 acoustic stimuli: (a) natural male 50-kHz USV and (b) time- and amplitude-matched white noise, with the latter serving as acoustic control for novelty-induced changes in behavior not linked to the communicative function of male prosocial 50-kHz USV. Our present findings show that female rats display high levels of social approach behavior in response to male prosocial 50-kHz USV, but not time- and amplitude-matched white noise, supporting the conclusion that male prosocial 50-kHz USV are likely to play an important role in establishing social proximity and possibly regulate mating behavior.