Ultrasonic vocalization by rat pups during recovery from deep hypothermia.

Vocalization in the ultrasonic range (USV) has been reported to occur in young rodents in response to isolation, novelty, handling, and cold. Heretofore these calls have been known to occur only in alert, attentive, or emotionally aroused animals. These studies describe the emission of USV by comatose 9- to 10-day-old rat pups during recovery from deep hypothermia. Calling began at 15-18 degrees C core temperature while pups were virtually unresponsive to stimulation. Experimental results describe the patterns of call production in relation to respiration, cardiac function, colonic temperature, and brown adipose tissue thermogenesis. These vocalizations were 32-42 kHz in frequency, reached peak rates of 50/min at 23 degrees C, and were eliminated by laryngeal denervation, thus resembling isolation-induced vocalizations. However, contact with their dams failed to reduce call rates until pups had warmed above 25 degrees C. Newborn and weanling pups also emitted USV in deep hypothermia, but no USV were observed in pups recovering from general anesthesia. The possible functions and evolution of this behavior are discussed.     

Potentiation and inhibition of ultrasonic vocalization of rat pups: Regulation by social cues

Eight- and 11- to 12-day-old rat pups were tested in isolation and in the presence of an anesthetized adult under cold conditions. Pups of both ages reduced rates of ultrasonic vocalization (USV) when an adult was placed into the test cage, independent of whether the adult was their dam or an unrelated male. However, afer removal of the dam, pups greatly increased their rates of USV over their first isolation period and in comparison with control pups. USV rates remained low after removal of the male. The temperature challenges faced by the pups in the two experimental conditions were the same. These phenomena are better explained by a hypothesis that postulates USV rate as being multiply determined, including by social cues, rather than a theory that considers thermal challenge only. © 1997 John Wiley & Sons, Inc. Dev Psychobiol 30: 195–200, 1997     

Rat pups reduce ultrasonic vocalization after exposure to an adult male rat

We examined how the experience of a threatening stimulus alters subsequent behavior in a situation where the immediate threat is absent. A small huddle of 12-day-old rats was exposed to a potentially infanticidal adult male rat for 5 min. During male exposure, pups were significantly more immobile than control pups. Thirty, 60, and 180 min after male exposure, the pups were isolated for 5 min from litter and dam in an unfamiliar environment. When isolated, pups that had been previously exposed to the male emitted significantly fewer ultrasonic vocalizations than controls, but did not differ in immobility. Low levels of vocalization were apparent 30 and 60 min after male exposure and were not evident at 180 min. The pups seemed to have adjusted their behavior to a potential male threat in a different context for a limited period of time.     

Isolation-induced vocalization in Wistar rat pups is not increased by naltrexone.

Rat pups, when removed from dam and littermates and isolated in an unfamiliar milieu, emit a characteristic ultrasonic vocalization that can be quieted by the administration of a nonsedating dose of morphine. Wistar pups, aged 7, 10, 12, 14, or 16 days, were tested after receiving intraperitoneal injections of the opiate antagonist naltrexone (0.0, 0.5, 1.0, or 5.0 mg/kg). The rate of isolation-induced ultrasonic vocalizations was unaffected by naltrexone at any dose, and there were no significant naltrexone-related changes on other behavioral measures. The complexity of the opioid system is discussed, as it may be involved in the vocal reaction to isolation.     

The influence of olfaction on potentiation and inhibition of ultrasonic vocalization of rat pups

Twelve-day-old isolated rat pups reduce their rates of ultrasonic vocalization (USV) when an anesthetized adult is placed into the test cage, whether the adult is their dam or an unfamiliar male. USV rates remain very low even after removal of the male (inhibition). However, after removal of the dam, pups greatly increase their rates of USV over their first isolation period (potentiation). USV potentiation can be induced by either an awake, normally behaving dam or by one that is anesthetized. To test the role of olfaction in inhibition and potentiation, PND12 pups were rendered anosmic via intranasal infusion of 5% zinc sulphate (ZnSO4). Control pups were infused with normal saline. After overnight separation from the dam, the USV and other behaviors of pups were recorded during a 6-min test. Each pup was tested during an initial isolation period and a final isolation period. In the first experiment, an anesthetized adult (dam or unfamiliar male) or no companion was placed in contact with the pup during the middle minute. Anosmia prevented both potentiation and inhibition of USV by passive adult contact. Thus, it seems likely that pups use olfactory discrimination as the basis for these two highly differentiated vocal responses to social stimuli. Results from two additional experiments demonstrate that anosmia does not prevent potentiation when the adult dam is active and interacting with the pup on either postnatal day 12 or 8.     

Repeated exposure of rat pups to isolation attenuates isolation-induced ultrasonic vocalization rates: Reversal with naltrexone

Young rat pups are dependent on the dam for their survival, thus isolation of the neonatal rat pup from the dam presents the young organism with a variety of stressors. The question examined in this study concerns the ability of the young rat pup to modify its response to isolation following repeated exposure to that isolation as well as the role played by endogenous opiates in this process. Following repeated isolations, pups were seen to decrease vocalization rates. Altering the context in an attempt to dishabituate animals failed to reverse the decreased vocalization rate. However, opiate receptor blockade attenuated this decrease when administered subsequent to the first isolation period but not prior to the last isolation period. These results suggest that the development of this attenuated response to isolation stress is opiate-mediated but that once established, its expression is not dependent on endogenous opiate release.© 1994 John Wiley & Sons, Inc.     

A cholecystokinin receptor antagonist blocks milk-induced but not maternal-contact-induced decrease of ultrasonic vocalization in rat pups

The role of cholecystokinin (CCK) in reducing separation-induced ultrasonic vocalization (USV) was examined by peripheral administration (of the selective CCK(A) receptor antagonist devazepide to 10-11-day-old rats. Pups placed alone for 2 min emitted a mean of 55.1 USV/min. When placed on a paper towel wet with warm, sweet milk, USV rate decreased to 23.2/min for the following 8 min. Devazepide (150-600 microg/kg IP) prevented this USV reduction, but did not increase feeding. In contrast, USV reduction produced by contact with the anesthetized dam was not affected by devazepide. Similarly, the opiate antagonist naltrexone (0.5 and 1.0 mg/kg) has been shown to block morphine-induced USV decrease in pups away from the dam, but was ineffective when USV reduction was induced by the presence of the dam (Blass et al., 1990; Carden & Hofer, 1990). The current findings suggest that CCK's role is specific, in that it mediates milk- but not dam-induced quieting of USV. The results, however, are not incompatible with the possibility that CCK and opioids are part of multiple, redundant pathways that mediate the quieting of USV by the dam.     

Perinatal high-fat diet impairs pup retrieval and induces sex-specific changes in ultrasonic vocalization characteristics of rat pups

Rodent pups emit ultrasonic vocalizations (USVs) to solicit maternal behavior, promoting their survival. Conversely, maternal behaviors affect the expression of pup USVs. We previously demonstrated that a maternal diet high in saturated fat (HFD) alters maternal behavior and is associated with early maturation of pups and their stress physiology. Here, we assessed the developmental profiles of pup USVs using quantitative and qualitative measures on postnatal days (PND)7 and 13. Quantitative measures included call counts, duration, and frequency, while qualitative measures examined calls' sonographic structures. HFD offspring lacked the typical decrease in USV numbers with age observed among control offspring. They also had shorter calls at PND7 compared to control and HFD offspring at PND13. HFD female offspring showed a greater number of one-frequency-sweep calls, while male pups showed a greater number of two-frequency-sweep calls compared to control offspring. Concomitantly, HFD dams showed impaired pup retrieval on PND7. The data suggest that fewer USVs of shorter duration in HFD offspring may alter dam solicitation and thus impair maternal pup retrieval. This study highlights the impacts of perinatal HFD exposure on the dyadic reciprocal interaction between dam and pups, which may set the stage for long-lasting effects on offspring physiology and behavior.     

Potentiation of isolation-induced vocalization by brief exposure of rat pups to maternal cues

Since their discovery in 1956, the highest rates of ultrasonic vocalization (USV) have been recorded from infant rats when first isolated in an unfamiliar place. We now report that peak USV rates can be doubled by allowing test pups a brief initial period of contact with their anesthetized dam (1–10 min) in the test chamber before isolating the pup by her removal. Potentiation of the isolation response was specific to the dam, for it failed to occur following initial contact with a group of 4 warm, anesthetized littermates. Control experiments showed that potentiation could not be attributed to thermal contrast, experimenter handling, general behavioral activation, novelty of maternal cues, or nursing deprivation. Furthermore, it did not occur when pups were taken for isolation testing directly from prolonged contact with their anesthetized dam in the home cage. Potentiation may be understood in terms of the communicative role of the pups' call and/or prior learning contingencies within the mother–infant interaction.©1994 John Wiley & Sons, Inc.     

Startle-inducing acoustic stimuli evoke ultrasonic vocalization in the rat

The present study demonstrates that acoustic stimuli which induce a startle response (ASR) also evoke ultrasonic vocalization in the rat. Sound recordings were done on three consecutive days of testing during sessions of 20 acoustic stimuli each and on the following day for three minutes following 5 acoustic stimuli (nonstimulus condition). Startle-inducing stimuli evoked continuous ultrasonic calling which was maintained throughout testing. Immediately following each acoustic stimulus, however, vocalization was interrupted by a period of silence (gap). The mean duration of sounds was reduced and the interpulse interval tended to increase during acoustic stimulation as compared to the nonstimulus condition. It is concluded that startle-eliciting stimuli induce a state of fear in the rat and that the acoustic-startle-elicited ultrasonic vocalization may provide a novel model in the study of anxiety.     

Ontogeny of noradrenergic effects on ultrasonic vocalizations in rat pups

The ontogeny of noradrenergic effects and the interaction of opioid and noradrenergic systems on vocalizations in rat pups from Day 10 to Day 18 were evaluated. Day 10 pups given clonidine (0.05 or 0.5 mg/kg) ip showed a sustained high level of calling throughout a 25-min isolation period that was reversed with yohimbine (0.1 mg/kg). Day 15 pups showed identical profiles with a lower baseline rate. Day 17 pups' calls were differentially affected according to dose; Day 18 pups reduced vocalizing with clonidine. In addition, it was found that at all ages when clonidine increased calling during isolation, the pups vocalized in the nest as well. Naltrexone, an opioid antagonist, lost its effectiveness to increase vocalizations after Day 15 unless it was given subsequent to clonidine. These results suggest that pups' vocalizations are differentially affected by noradrenergic and opioid stimulation or inhibition with developmental changes.     

Thermogenesis and thermolysis during sleeping and waking in the rat

Thermogenesis (VO₂), sensible heat loss and subcutaneous back temperature were recorded simultaneously during sleeping and waking in both intact and depilated rats at Ta ranging from 21–28°C.VO₂ increased during wakefulness (W), decreased and plateaued during slow wave sleep (SWS) and then decreased 10% with each paradoxical sleep (PS) phase. Sensible heat loss, which represented about 90% of the heat production, increased and plateaued during SWS, decreased in W and generally rose abruptly (+40%) during PS. After removal of the fur the mean levels ofVO₂ and sensible heat loss were increased by 30–50% and returned to normal values within two weeks, although their variations related to stages of sleep were unchanged.These results concerning thermogenesis and thermolysis are in agreement with the variations of body temperature (brain excluded) during sleeping and waking.     

Prenatal stress suppresses rat pup ultrasonic vocalization and myoclonic twitching in response to separation

Prenatally stressed infant rats were separated from their dams and littermates on postnatal Day 14 and their rates of ultrasonic vocalizations (USVs) and myoclonic twitching in response to that separation were recorded. Compared to control pups, prenatally stressed pups vocalized significantly less often and showed significantly less myoclonic twitching in response to this test. Results are interpreted in terms of the ability of prenatal stress to produce infants that are behaviorally inhibited when in a novel or stressful situation.     

Effect of a social companion on the ultrasonic vocalizations and contact responses of 3-day-old rat pups.

Rat pups that are isolated in a novel environment emit ultrasonic calls. Vocalization in 3-day-old pups has been thought to be predominantly under thermal control (Allin & Banks, 1971). By the 2nd week of life, ultrasonic vocalizations are reduced when pups are tested in the company of a single anesthetized littermate (Carden & Hofer, 1990a; Hofer & Shair, 1978). In the present study, it was demonstrated that the vocalization rate of 3-day-old pups is also decreased when an anesthetized littermate is present in the isolation chamber. To determine whether this quieting was a function of the body heat of the companion, in a 2nd experiment, the axillary temperature of the companion was lowered until it was the same as the ambient temperature of the test chamber, 22 degrees C. In the presence of a cool companion, ultrasonic vocalizations were also reduced. For the last experiment, a textured plastic surrogate was substituted for the littermate. Calls were not diminished in the presence of the surrogate.     

Control of ingestion by oral habituation in rat pups.

The role of habituation of mouthing activity in the control of ingestion was investigated in 6-, 12-, and 18-day-old rat pups. In pups at all ages, oral habituation to a flavored diet inhibited ingestion of a continuous oral infusion of that same diet. Twelve-day-old pups that had orally habituated to a diet continued to consume less of a continuous oral infusion of that diet both 30 min and 3 hr later, and the duration of suppressed ingestion was shown to be dependent on the rate of stimulus presentation during habituation experience. These data suggest that oral habituation may be a diet-specific influence on both intra- and intermeal patterning.     

Dopaminergic modulation of kappa opioid-mediated ultrasonic vocalization, antinociception, and locomotor activity in the preweanling rat.

The purpose of this study was to determine whether dopamine (DA) systems modulate kappa opioid-mediated ultrasonic vocalizations (USVs), antinociception, and locomotion in young rats. Seventeen-day-old rats were injected with the kappa agonist U-50,488 (0.0-7.5 mg/kg) and saline, the D2-like receptor agonist R(-)-propylnorapomorphine (NPA; 0.1 or 1.0 mg/kg), the indirect DA agonist cocaine (10 or 20 mg/kg), or the DA antagonist flupenthixol (0.25 or 0.5 mg/kg). USVs and locomotion were measured for 6 min, with antinociception being assessed with a tail-flick test. Kappa receptor stimulation produced analgesia and increased USVs and locomotion. U-50,488-induced analgesia was potentiated by NPA, whereas U-50,488-induced USVs were attenuated by both DA agonists. NPA and flupenthixol depressed U-50,488's locomotor effects. These results show that DA systems interact with kappa opioid systems to modulate USVs, antinociception, and locomotion in preweanling rats.     

Developmental changes in the suckling behavior of hamster pups: a comparison with rat pups

The suckling behavior of hamster pups was observed to undergo several transitions during the preweaning period. Pups 10 days of age and younger attached to the nipples of their anesthetized mothers and suckled. After 10 days of age, however, only animals which had been deprived of suckling (and food and water) reliably attached to their mothers' nipples. By 25 days of age, deprived pups no longer suckled. The pattern of response after nipple attachment also changed during development: pups younger than 5 days of age remained on 1 nipple after initial attachment, whereas older pups shifted from 1 nipple to another. The development of suckling in hamster pups resembles that in rat pups, although certain changes or transitions occur at different ages in the 2 species. These changes may provide important markers for neural and behavioral development in each species and for comparisons between the two.     

Nutritional variables and their effect on the development of ultrasonic vocalizations in rat pups

Measures of ultrasonic vocalizations were made on alternate days during the first and second weeks postpartum in rat pups reared by mothers fed either control (23%) or low protein (8%) diets. Compared to those reared by control mothers, pups reared by low protein mothers emitted significantly fewer ultrasonic vocalizations. The hypothesis that malnutrition might shift the age at which peak rate of vocalizations were emitted was not confirmed.     

Development of ultrasonic calls in rat pups follows similar patterns regardless of isolation distress

Ultrasonic vocalizations (USVs) emitted by rat pups have been extensively studied in the context of isolation from their mother and littermates. The heightened call rate of isolation-induced USVs indicates increased anxiety, whereas other acoustic parameters, such as frequency and call duration, are thought to be useful in reflecting developmental changes, which by weaning have developed into their adult form. However, it is possible that stress can affect the quality as well as the quantity of calls, and that as the pups become more mobile, the effects of the stress may change. Therefore, in the present study rats were tested in a test arena either in isolation or with littermates, so as to assess the effects of isolation stress on the developmental changes reported in the literature. Rat pups of both sexes were tested on multiple days from postnatal day (PND) 7 to weaning. Overall, the main changes in the frequency, duration, and types of calls were accounted for by age. The effects of isolation were minor, as were sex differences. This study indicates that the development of USVs in infant rats is a robust process and seemingly resistant to the effects of isolation-induced stress.     

Gastric saline infusion reduces ultrasonic vocalizations and brown fat activity in suckling rat pups

Under standard conditions involving isolation and cooling, it has been documented that intraoral infusion of milk and injection of the intestinal peptide cholecystokinin (CCK) result in an attenuation in ultrasonic vocalizations (USV) emitted by infant rat pups. One of the most effective stimuli in inhibiting ingestion in suckling rat pups is gastric distension, but the effect of gastric distension on USV production has not been reported. In this experiment, we subjected infant rats to intragastric infusion of isotonic saline (2% body weight) to produce a natural level of gastric distension and hydration. We found that this stimulus resulted in a powerful reduction in USV emissions in isolated 10-day-old rats. In a subsequent experiment, we found that gastric saline infusion also diminished brown adipose tissue (BAT) thermogenesis. There were different time courses of the gastric saline infusion effects on BAT thermogenesis and on USV emissions, however, suggesting that these processes may be independently regulated. We hypothesize that this stimulus induces a transient activation of the parasympathetic nervous system, which overrides the sympathetic control of BAT and USV production.