Rat Ultrasonic Vocalizations Research Articles

Vocalization-whisking coordination and multisensory integration of social signals in rat auditory cortex.

Social interactions involve multi-modal signaling. Here, we study interacting rats to investigate audio-haptic coordination and multisensory integration in the auditory cortex. We find that facial touch is associated with an increased rate of ultrasonic vocalizations, which are emitted at the whisking rate (∼8 Hz) and preferentially initiated in the retraction phase of whisking. In a small subset of auditory cortex regular-spiking neurons, we observed excitatory and heterogeneous responses to ultrasonic vocalizations. Most fast-spiking neurons showed a stronger response to calls. Interestingly, facial touch-induced inhibition in the primary auditory cortex and off-responses after termination of touch were twofold stronger than responses to vocalizations. Further, touch modulated the responsiveness of auditory cortex neurons to ultrasonic vocalizations. In summary, facial touch during social interactions involves precisely orchestrated calling-whisking patterns. While ultrasonic vocalizations elicited a rather weak population response from the regular spikers, the modulation of neuronal responses by facial touch was remarkably strong.

Critical involvement of 5-HT2C receptor function in amphetamine-induced 50-kHz ultrasonic vocalizations in rats.

Rats emit various distinct types of ultrasonic vocalizations (USV), with high-frequency 50-kHz USV typically occurring in appetitive situations being elicited by administering drugs of abuse, most notably amphetamine (AMPH), possibly reflecting drug wanting/craving and/or liking. Because 50-kHz USV emission is, at least in part, dopamine (DA) dependent and 5-HT2C agonists inhibit DA neurotransmission, we hypothesized that AMPH-induced 50-kHz USV can be attenuated by pretreatment with a 5-HT2C agonist. In experiments I and II, a dose-response curve for AMPH-induced 50-kHz USV was established, and the partial dependency of AMPH-induced 50-kHz USV on DA neurotransmission was validated by pretreatment with the D2-antagonist eticlopride. In experiment III, rats were pretreated with the 5-HT2C agonist CP 809,101 (0.0, 0.3, 1.0, 3.0, and 10 mg/kg), while in experiment IV, CP 809,101 (3.0 mg/kg), the 5-HT2C antagonist SB 242084 (1.0 mg/kg), or the combination of the two, was applied before AMPH administration (2.0 mg/kg). Finally, in experiment V, rats were treated with SB 242084 (0.0, 0.1, 0.3, and 1.0 mg/kg) only, i.e., in absence of AMPH. The 5-HT2C agonist CP 809,101 dose-dependently blocked AMPH-induced 50-kHz USV, most notably trills, a call subtype that is considered to exclusively reflect a positive affective state, while the 5-HT2C antagonist SB 242084 induced opposite effects. Moreover, SB 242084 induced 50-kHz USV by its own. 5-HT2C receptors are critically involved in AMPH-induced 50-kHz USV, with 5-HT2C antagonism resulting in a stimulant-like effect. Attenuation of drug wanting/craving and/or liking by coadministration of a 5-HT2C agonist could be a translational pharmacodynamic biomarker.

Rodent ultrasonic vocalizations are bound to active sniffing behavior.

During rodent active behavior, multiple orofacial sensorimotor behaviors, including sniffing and whisking, display rhythmicity in the theta range (~5–10 Hz). During specific behaviors, these rhythmic patterns interlock, such that execution of individual motor programs becomes dependent on the state of the others. Here we performed simultaneous recordings of the respiratory cycle and ultrasonic vocalization emission by adult rats and mice in social settings. We used automated analysis to examine the relationship between breathing patterns and vocalization over long time periods. Rat ultrasonic vocalizations (USVs, “50 kHz”) were emitted within stretches of active sniffing (5–10 Hz) and were largely absent during periods of passive breathing (1–4 Hz). Because ultrasound was tightly linked to the exhalation phase, the sniffing cycle segmented vocal production into discrete calls and imposed its theta rhythmicity on their timing. In turn, calls briefly prolonged exhalations, causing an immediate drop in sniffing rate. Similar results were obtained in mice. Our results show that ultrasonic vocalizations are an integral part of the rhythmic orofacial behavioral ensemble. This complex behavioral program is thus involved not only in active sensing but also in the temporal structuring of social communication signals. Many other social signals of mammals, including monkey calls and human speech, show structure in the theta range. Our work points to a mechanism for such structuring in rodent ultrasonic vocalizations.

Behavioral Correlates of 50-kHz Ultrasonic Vocalizations in Rats: Progressive Operant Discrimination Learning Reduces Frequency Modulation and Increases Overall Amplitude

Ultrasonic vocalizations (USVs) in rats are thought to contain ecological signals reflecting emotional states. These USVs are centered on 50-kHz, and frequency modulation (FM) is hypothesized to indicate positive emotion; however, results from recent studies are inconsistent with this hypothesis. We suspected that such inconsistencies might result from ambiguity in defining frequency modulation, and problems with acoustic analyses and behavioral protocols. We addressed these problems by applying quantitative methods for USV analyses and using a food reward operant paradigm. Our results revealed that frequency modulation varied according to the degree of positive outcomes, but the direction of change was opposite to what had been observed in previous studies. The FM in 50-kHz USVs decreased as animals learned the task and obtained more reinforcement, while USV amplitude increased as learning progressed. To reconcile these results with those from prior studies, we suggest that FM in 50-kHz USVs should be taken as an index of reward prediction errors, and USV amplitude should be considered as an index of positive emotion.

Emotion and relative reward processing: An investigation on instrumental successive negative contrast and ultrasonic vocalizations in the rat

Incentive contrast effects include changes in behavioral responses after a reward upshift (positive contrast) or downshift (negative contrast). Proposed influences on these behavioral changes are emotional state reactions after experiencing or anticipating a change in reward outcome. Rat ultrasonic vocalizations have been shown to be indicators of emotional state during behavior and anticipatory periods. The objective of the present study was to monitor rodent ultrasounds during incentive contrast using a classical runway procedure called instrumental successive negative contrast. The procedure is one that has been used often to examine incentive relativity because of its reliability in measuring negative contrast effects. Rats were trained to run in the alleyway to receive a high (12 pellets) or low magnitude (1 pellet) outcome. The high magnitude was then shifted to the low and running speeds in the alleyway for the reward and USV emission were compared. Replicating previous work, a negative contrast effect was observed with postshift running speeds significantly slower in the shifted group compared to the unshifted group. USVs did not follow the same pattern with an apparent lack of significant differences between the groups following the reward downshift. We also tested another group of animals using a visual predictive cue in the same runway test. When visual cues predicted high or low magnitude outcome, no incentive contrast was found for the running speeds following an outcome downshift, but a weak contrast effect was observed for the USV emission. These results demonstrate a separation between USVs and behavioral indicators of incentive contrast suggesting that concomitant shifts in negative affect may not be necessary for anticipatory relative reward processes.

Phasic dopamine release in the nucleus accumbens in response to pro-social 50 kHz ultrasonic vocalizations in rats.

Rats emit ultrasonic vocalizations (USVs) that are thought to serve as situation-dependent affective signals and accomplish important communicative functions. In appetitive situations, rats produce 50 kHz USVs, whereas 22 kHz USVs occur in aversive situations. Reception of 50 kHz USVs induces social approach behavior, while 22 kHz USVs lead to freezing behavior. These opposite behavioral responses are paralleled by distinct brain activation patterns, with 50 kHz USVs, but not 22 kHz USVs, activating neurons in the nucleus accumbens (NAcc). The NAcc mediates appetitive behavior and is critically modulated by dopaminergic afferents that are known to encode the value of reward. Therefore, we hypothesized that 50 kHz USVs, but not 22 kHz USVs, elicit NAcc dopamine release. While recording dopamine signaling with fast-scan cyclic voltammetry, freely moving rats were exposed to playback of four acoustic stimuli via an ultrasonic speaker in random order: (1) 50 kHz USVs, (2) 22 kHz USVs, (3) time- and amplitude-matched white noise, and (4) background noise. Only presentation of 50 kHz USVs induced phasic dopamine release and elicited approach behavior toward the speaker. Both of these effects, neurochemical and behavioral, were most pronounced during initial playback, but then declined rapidly with subsequent presentations, indicating a close temporal relationship between the two measures. Moreover, the magnitudes of these effects during initial playback were significantly correlated. Collectively, our findings show that NAcc dopamine release encodes pro-social 50 kHz USVs, but not alarming 22 kHz USVs. Thus, our results support the hypothesis that these call types are processed in distinct neuroanatomical regions and establish a functional link between pro-social communicative signals and reward-related neurotransmission.

Attribution and expression of incentive salience are differentially signaled by ultrasonic vocalizations in rats.

During Pavlovian incentive learning, the affective properties of rewards are thought to be transferred to their predicting cues. However, how rewards are represented emotionally in animals is widely unknown. This study sought to determine whether 50-kHz ultrasonic vocalizations (USVs) in rats may signal such a state of incentive motivation to natural, nutritional rewards. To this end, rats learned to anticipate food rewards and, across experiments, the current physiological state (deprived vs. sated), the type of learning mechanism recruited (Pavlovian vs. instrumental), the hedonic properties of UCS (low vs. high palatable food), and the availability of food reward (continued vs. discontinued) were manipulated. Overall, we found that reward-cues elicited 50-kHz calls as they were signaling a putative affective state indicative of incentive motivation in the rat. Attribution and expression of incentive salience, however, seemed not to be an unified process, and could be teased apart in two different ways: 1) under high motivational state (i.e., hunger), the attribution of incentive salience to cues occurred without being expressed at the USVs level, if reward expectations were higher than the outcome; 2) in all experiments when food rewards were devalued by satiation, reward cues were still able to elicit USVs and conditioned anticipatory activity although reward seeking and consumption were drastically weakened. Our results suggest that rats are capable of representing rewards emotionally beyond apparent, immediate physiological demands. These findings may have translational potential in uncovering mechanisms underlying aberrant and persistent motivation as observed in drug addiction, gambling, and eating disorders.

Rat ultrasonic vocalization shows features of a modular behavior.

Small units of production, or modules, can be effective building blocks of more complex motor behaviors. Recording underlying movements of vocal production in awake and spontaneously behaving male Sprague Dawley rats interacting with a female, I tested whether the underlying movements of ultrasonic calls can be described by modules. Movements were quantified by laryngeal muscle EMG activity and subglottal pressure changes. A module was defined by uniformity in both larynx movement and pressure pattern that resulted in a specific spectrographic feature. Modules are produced either singly (single module call type) or in combination with a different module (composite call type). Distinct modules were shown to be linearly (re)combined. Additionally, I found that modules produced during the same expiratory phase can be linked with or without a pause in laryngeal activity, the latter creating the spectrographic appearance of two separate calls. Results suggest that combining discrete modules facilitates generation of higher-order patterns, thereby increasing overall complexity of the vocal repertoire. With additional study, modularity and flexible laryngeal-respiratory coordination may prove to be a basal feature of mammalian vocal motor control.

The incentive value of males’ 50-kHz ultrasonic vocalizations for female rats (Rattus norvegicus).

Rats emit ultrasonic vocalizations (USVs) of 22 kHz and 50 kHz before, during, and after copulation. The 50-kHz vocalizations can be subdivided into flat and frequency-modulated (FM) trill calls. In this study, the role of 50-kHz USVs in sexual incentive motivation in female rats was examined. USVs were recorded from sexually active males during the precopulatory phase. In the first two experiments, "full 10-min song," flat-call, and FM-trill-call stimuli were used. In Experiment 1, a combination of complex and trill calls was used as the FM-trill call, whereas a mixture of multistep and upward ramp calls was used in Experiment 2. The auditory stimuli were played back to sexually receptive female rats in a sexual incentive-motivation test. For comparison, the odor of an intact male rat was also used as incentive stimulus. The flat-call stimulus did not induce approach behavior in any experiment, whereas the FM trill showed a short-lived effect in Experiment 2. The females approached the "full song" during the first minute of stimulus exposure in Experiment 1, but not in Experiment 2. When the entire 10-min test period was analyzed, the females in Experiment 1 did not approach the full song more than background noise, though they did so in Experiment 2. The olfactory stimulus, to the contrary, had a clear incentive value in both experiments. In a third experiment, a devocalized male, an intact vocalizing male and a female rat were used as incentive stimuli. The females did not approach a vocalizing male more than they approached a silent male, showing that USVs do not contribute to the male rat's incentive value. Overall, the results of the present series of experiments show that the male rat's USVs do not have any consistent incentive value for the sexually receptive female rat. This sharply contrasts the strong and reliable effect of male odor.

Acute, but not repeated, administration of the neurotensin NTS1 receptor agonist PD149163 decreases conditioned footshock-induced ultrasonic vocalizations in rats

Neurotensin is an endogenous neuropeptide that has significant interactions with monoamine neurotransmitter systems. To date, neurotensin NTS1 receptor agonists, such as PD149163, have been primarily evaluated for the treatment for schizophrenia, drug addiction, and pain. Recently, PD149163 was found to attenuate fear-potentiated startle in rats, an experimental procedure used for screening anxiolytic drugs. The present study sought to assess these findings through testing PD149163 in a conditioned footshock-induced ultrasonic vocalization (USV) model. Conditioning was conducted in male Wistar rats using chambers equipped with shock grid floors and an ultrasonic vocalization detector. PD149163 and the 5-HT1A receptor partial agonist buspirone produced a statistically significant reduction of 22kHz USV counts. The typical antipsychotic haloperidol also reduced 22kHz USV counts, but did so at cataleptic doses. Ten days of repeated administration of PD149163 abolished the inhibitory effects of PD149163 on 22kHz USVs. These findings further support an anxiolytic profile for PD149163. However, tolerance to these effects may limit the utility of these drugs for the treatment of anxiety.

Dopamine D1 and D2 receptor antagonism effects on rat ultrasonic vocalizations

Voice disorders manifest in the early stages of Parkinson disease (PD), suggesting the vulnerability of the laryngeal sensorimotor system to mild alterations in dopamine signaling. Previous research has demonstrated that manipulations of central dopamine result in acoustic changes in rat ultrasonic vocalization (USV) and selective manipulation of receptor subtypes results in dose dependent changes in call rate and complexity. However, no study has specifically focused on the influence of dopamine receptor subtypes on acoustic features of USV production. This study examined the influence of D1 and D2 receptor subtypes on voluntary laryngeal sensorimotor control (USV) and gross whole-body involvement. Rat USV acoustics and catalepsy descent time were analyzed following the administration of selective D1 and D2 receptor antagonists in isolation and in combination, and a vehicle control. Results support the hypothesis that degradations of the acoustic signal would be most severe following combined receptor antagonism (D1+D2) compared with D1 or D2 receptor antagonism alone, and the vehicle (saline) condition. In addition, results indicate that selective D1 receptor antagonism alters acoustic parameters to a greater extent than D2 receptor antagonism. Thus, dopamine receptor subtypes appear to influence acoustic parameters to different degrees. Catalepsy descent time was longest following combined dopamine receptor antagonism but was also significantly increased with selective D1 or D2 antagonism. Together, these results support the potentially different contributions receptor subtypes play in cranial and limb sensorimotor control.

Vocal Training Mitigates Age-Related Changes Within the Vocal Mechanism in Old Rats

Aging affects voice production and is associated with reduced communicative ability and quality of life. Voice therapy is a critical component of treatment, but its effects on neuromuscular mechanisms are unknown. The ultrasonic vocalizations (USVs) of rats can be used to test the effects of aging and voice use on the laryngeal neuromuscular system. This study tested the hypothesis that age-related changes in the USVs of rats and laryngeal neuromuscular junctions can be reversed through vocal exercise. Young and old rats were trained for 8 weeks to increase their USVs and were compared with a no intervention group pre- and post-treatment. USV acoustics and aspects of neuromuscular junction (NMJ) morphology were measured in the thyroarytenoid muscle. Vocal training reduced or eliminated some age differences found in both USVs and NMJs. We conclude that vocal exercise may assist in mitigating age-related changes in voice characteristics and underlying neuromuscular adaptations.

Rat ultrasonic vocalizations demonstrate that the motivation to contextually reinstate cocaine‐seeking behavior does not necessarily involve a hedonic response

Human self-reports often indicate that changes in mood are a major contributor to drug relapse. Still, arguments have been made that instances of drug-seeking following abstinence in animal models (i.e. relapse/reinstatement) may be outside of hedonic control. Therefore, the present study utilized ultrasonic vocalizations in the rat in order to evaluate affect during cocaine self-administration and contextual reinstatement of cocaine-seeking in a pre-clinical model of drug relapse (abstinence-reinstatement model). Results show that while subjects effectively reinstated drug-seeking (lever pressing) following 30 days of abstinence, and spontaneously recovered/reinstated drug-seeking following 60 days of abstinence, ultrasonic vocalizations did not increase over baseline levels during either reinstatement session. These results are consistent with previous results from our laboratory and current theories of addiction suggesting that cues that are weakly associated with drug consumption can motivate drug-seeking behavior that is outside of hedonic processing.

Levomilnacipran (F2695), a norepinephrine-preferring SNRI: Profile in vitro and in models of depression and anxiety

Levomilnacipran (LVM; F2695) is the more active enantiomer of the serotonin/norepinephrine (5-HT/NE) reuptake inhibitor (SNRI) milnacipran and is currently under development for the treatment of major depressive disorder. LVM was benchmarked against two other SNRIs, duloxetine and venlafaxine, in biochemical, neurochemical and pharmacological assays. LVM exhibited high affinity for human NE (Ki = 92.2 nM) and 5-HT (11.2 nM) transporters, and potently inhibited NE (IC50 = 10.5 nM) and 5-HT (19.0 nM) reuptake (human transporter) in vitro. LVM had 2-fold greater potency for norepinephrine relative to serotonin reuptake inhibition (i.e. NE/5-HT potency ratio: 0.6) and 17 and 27 times higher selectivity for NE reuptake inhibition compared with venlafaxine and duloxetine, respectively. LVM did not exhibit affinity for 23 off-target receptors. LVM (i.p.) increased cortical extracellular levels of 5-HT, and NE (minimal effective doses: MEDs = 20 and 10 mg/kg, respectively). In anti-depressive/anti-stress models, i.p. LVM diminished immobility time in the mouse forced swim (MED = 20 mg/kg) and tail suspension (MED = 2.5 mg/kg) tests, and reduced shock-induced ultrasonic vocalizations in rats (MED = 5 mg/kg). Duloxetine and venlafaxine were less potent (MEDs ≥ 10 mg/kg). At doses active in these three therapeutically-relevant models, LVM (i.p.) did not significantly affect spontaneous locomotor activity. In summary, LVM is a potent, selective inhibitor of NE and 5-HT transporters with preferential activity at the former. It is efficacious in models of anti-depressive/anti-stress activity, with minimal potential for locomotor side effects.

CALL FOR INTERACTION OF ULTRASOUND IN RATS

The article presents data illustrating the changes in the characteristics of ultrasonic vocalizations in rats (mean frequency, duration and power ranges of ultrasonic waves) in acute emotional stressor impact. In the ultrasonic vocalizations of rats reflects individual motivation emotive, dominating at a certain time. These data illustrate the importance of ultrasound information in mammals.

The role of dopaminergic transmission through D1-like and D2-like receptors in amphetamine-induced rat ultrasonic vocalizations

Systemic amphetamine (AMPH) administration increases the rate of 50-kHz ultrasonic vocalizations (USVs) in adult rats and preferentially enhances the 'trill' subtype; these effects of AMPH critically depend on noradrenergic transmission, but the possible contributions of dopamine are unclear. To assess the role of dopamine in 50-kHz USVs emitted drug-free and following systemic AMPH administration. Adult male Long-Evans rats pre-selected for high AMPH-induced calling rates were tested with AMPH (1 mg/kg, intraperitoneal (IP)) and saline following pretreatment with the following dopamine receptor antagonists: SCH 23390 (0.005-0.02 mg/kg, subcutaneous (SC)), SCH 39166 (0.03-0.3 mg/kg, SC), haloperidol (0.1, 0.2 mg/kg, IP), sulpiride (20-80 mg/kg, SC), raclopride (0.1-0.5 mg/kg, SC), clozapine (4 mg/kg, SC), risperidone (0.5 mg/kg, SC), and pimozide (1 mg/kg, IP). The dopamine and noradrenaline reuptake inhibitors (GBR 12909 and nisoxetine, respectively) were also tested, alone and in combination. SCH 23390, SCH 39166, haloperidol, and raclopride dose-dependently inhibited vocalizations under AMPH and suppressed the proportion of trill calls. Sulpiride, however, had no discernable effect on call rate or profile, even at a high dose that reduced locomotor activity. Single doses of clozapine, risperidone, and pimozide all markedly decreased calling under saline and AMPH. Finally, GBR 12909 and nisoxetine failed to promote 50-kHz USVs detectably or alter the subtype profile, when tested alone or in combination. The rate of 50-kHz USVs and the call subtype profile following systemic AMPH administration depends on dopaminergic neurotransmission through D1-like and D2-like receptors. However, inhibiting dopamine and/or noradrenaline reuptake appears insufficient to induce calling.

Effects of varying reinforcement probability on pavlovian approach behavior and ultrasonic vocalizations in rats

Long Evans rats (n=32) were trained for 2 weeks to respond to an auditory conditioned stimulus (CS) which signaled the delivery of a 20% sucrose unconditioned stimulus (US) with varying probabilities. Animals were randomly assigned to 1 of 4 groups. In the control groups, the CS signaled sucrose delivery with equal probabilities across two weeks, at 100% (Group 100-100) and 25% (Group 25-25) respectively. In the experimental groups (Group 100-25) and (Group 25-100), sucrose probabilities were switched between weeks 1 and 2. Three behavioral measures were recorded: latency to enter the sucrose port upon CS presentation, head entries throughout the session and ultrasonic vocalizations. The results suggest that all groups formed associations between the CS and US, as evidenced by a decrease in latency to respond to the CS across days. The experimental groups were also able to detect when sucrose probability changed, as evidenced by Group 25-100's increase in head entries, to the level of Group 100-100 in week 2, and Group 100-25's decrease in head entries, to the level of Group 25-25 in week 2. Group 100-25 also produced an increase in “22kHz” ultrasonic vocalizations following the downshift on the first day of week 2. The increase in this ultrasonic frequency range, which is associated with negative affect in rats, preceded both the decrease in head entries and the increase in missed trials, consistent with a multistage model of behaviors resulting from US probability reduction.

A rodent “self-report” measure of methamphetamine craving? Rat ultrasonic vocalizations during methamphetamine self-administration, extinction, and reinstatement

Rats emit ultrasonic vocalizations (USVs) in a variety of contexts, and it is increasingly clear that USVs reflect more complex information than mere positive and negative affect states. We sought to examine USVs in a common model of addiction and relapse, the self-administration/reinstatement paradigm, in order to gain insight into subjective states experienced by rats during various types of methamphetamine seeking. We measured three subtypes of “50kHz” USVs [flats, trills, and non-trill frequency modulated (FM) USVs], as well as long and short duration “22kHz” USVs, during self-administration and extinction training, and during reinstatement elicited by cues, a methamphetamine prime, cues+prime, or the pharmacological stressor yohimbine. During self-administration and extinction, rats emitted many flats and FMs, (and short duration “22kHz” USVs on day 1 of self-administration), but few trills. In contrast, methamphetamine priming injections potently enhanced FMs and trills, and trill production was correlated with the degree of methamphetamine+cue-elicited reinstatement. Cues alone yielded increases only in flat USVs during reinstatement, though a subset of rats displaying strong cue-induced reinstatement also emitted long duration, aversion-related “22kHz” USVs. Although yohimbine administration caused reinstatement, it did not induce “22kHz” USVs in methamphetamine-experienced or methamphetamine-naïve rats (unlike footshock stress, which did induce long duration “22kHz” USVs). These findings demonstrate heterogeneity of rat USVs emitted during different types of methamphetamine seeking, and highlight their potential usefulness for gaining insight into the subjective states of rats in rodent models of drug addiction and relapse.

Laryngeal aging and acoustic changes in male rat ultrasonic vocalizations

This study examined changes in the acoustic and temporal structure of ultrasonic vocalizations as a function of age and correlated acoustic changes with vocal fold microstructure. Ultrasonic vocalizations were recorded in three age groups of male rats: aged (24-26 months), middle-aged (17-18 months), and young (4-5 months). Acoustic and structural changes in vocal fold tissue were evident by 18 months of age. Histological analyses revealed decreased density of elastin and hyaluronic acid and increased collagen density in the middle-aged and aged groups compared to the young rats. Laryngeal microstructure correlated with some of the ultrasonic acoustic features. These results show that male Long Evans rats experience changes in ultrasonic acoustic structure by middle age, and these changes correlate with deterioration in laryngeal microstructure. Ultrasonic vocalizations can be used as a model system for age-related degeneration in vocal fold structure and function.

High-frequency ultrasonic vocalizations in rats in response to tickling: The effects of restraint stress

We tested the hypotheses that (a) the propensity to emit high-frequency (∼50kHz) ultrasonic vocalizations in response to manual “tickling” by an experimenter, may serve as a behavioral marker of positive affect in rats and, (b) that tickling may reduce the severity of stress. Group-housed adult rats were subjected to the 15-s tickling procedure daily, and their ultrasonic vocalization response was measured over a period of two weeks, until it has stabilized. The animals were then subjected to the restraint stress lasting for one week. The experimental groups were exposed to stress 1h before or 1h after tickling and the controls were tickled without stressing. Rats that were stressed 1h before tickling demonstrated a decreased number of the high-frequency calls as compared with the non-stressed controls. Stressing 23h before tickling reduced the call response less effectively. The propensity to emit high-frequency calls has normalized 7 and 12 days following the end of stressing. In addition, stressed groups showed a diminution of sucrose preference, which in the case of rats stressed 23h before tickling persisted even for 12 days following the end of restraint. The present data suggest that repeated stress may decrease the propensity to produce high-frequency vocalizations, and that this measure may serve as a biomarker of the depressive state of animals.