Thermal preference behavior in preweaning genetically obese (ob/ob) and lean (+/?, +/+) mice

Impaired nonshivering thermogenesis and lowered rectal temperatures (Tre) are hallmarks that appear early in the postnatal ontogeny of the genetically obese (ob/ob) mouse. Adult obese mice compensate behaviorally for these impairments and do not defend their low Tres. We predicted that, because young mice primarily rely on behavior to ensure thermal homeostasis during preweaning development, the appearance of the obese mouse's thermoregulatory impairment should promote their continued reliance on behavioral thermoregulation compared to lean pups. Accordingly, intact litters of pups from heterozygous lean (C57BL/6J, ob/+) and from homozygous lean (+/+) matings were tested at 6, 12, and 18 days postpartum on a thermal gradient (14-44 degrees C). Obese pups had lower pretest Tres than lean (+/?) littermates at 6 days and lower pretest Tres than both lean littermates and homozygous (+/+) lean control pups at 12 and 18 days. Exposure to the gradient ameliorated these differences (i.e., no posttest Tre differences among phenotypes). Correspondingly, obese pups preferred warmer gradient locations than +/+ pups but similar locations to their phenotypically lean (+/?) littermates until 18 days, when both lean groups preferred similar thermal locations compared to warmer-seeking obese pups. These data support our hypothesis and emphasize the age-dependent impact of the ob gene on altering mouse pups' thermal preferences.     

Competition and cooperation among huddling infant rats

Huddling is expressed by infant rats and continues to be an important behavior throughout adulthood. As a form of behavioral thermoregulation, huddling is thought to play an essential role in compensating for inadequate physiological thermoregulation early in development. Infant rats, however, are capable of heat production shortly after birth using brown adipose tissue (BAT) and exhibit thermogenesis in the huddle, suggesting that huddling does not obviate the need for endothermy during cold exposure. In the present experiment, 4-pup huddles of infant rats (2- or 8-day-olds) were exposed to two subthermoneutral temperatures, and BAT thermogenesis was inhibited in 0, 2, or 4 of the rats in each huddle. Inhibition of BAT thermogenesis compromised the pups' ability to maintain huddle temperature, but surprisingly did not result in enhanced huddling at either age. These results suggest that effective huddling during cold exposure requires the thermal resources provided by endothermy. Furthermore, the heat provided by BAT appears to shape behavioral interactions in the huddle during development.     

Independence of ultrasonic vocalization and thermogenic responses in infant rats

Ultrasonic vocalizations (USV) normally accompany brown adipose tissue (BAT) thermogenesis in infant rats exposed to cold. BAT activation (measured by implanted thermistors) was pharmacologically blocked with hexamethonium (20 mg/kg ip) in 12-13-day-old pups, but they nevertheless showed normal USV responses to cold. Activation of BAT in warm pups by norepinephrine (800 micrograms/kg sc) failed to elicit USV. It is concluded that BAT activation is neither necessary nor sufficient for USV production. To evaluate how tightly the two responses may be coupled centrally, rat pups deprived of nutrients for 24 hr, in which sympathetic activation is known to be inhibited centrally (Bignall, Heggeness, & Palmer, 1975), were studied. These pups vocalized with the same latency in response to cold as normals but failed to show evidence of concurrent BAT activation. It is concluded that USV and BAT thermogenesis are normally elicited together by cold but are not tightly linked physiologically.     

Evidence of a defect in energy expenditure in 7-day-old Zucker rat (fa/fa)

The aim of the investigation was to see whether a defect in energy expenditure could be found in the Zucker rat at the onset of obesity. Obese (fa/fa) and lean (Fa/fa) 7-day-old pups were studied at three ambient temperatures. At 33 degrees C fa/fa pups showed a reduction in oxygen consumption, respiratory CO2 production, in vivo oxidation of injected [1-14C]palmitic acid, as well as in core temperature. When the pups were kept at 28 degrees C, the difference between genotypes was considerably accentuated, thus indicating a subnormal thermogenic response of the fa/fa pups to a mildly cold environment. At 20 degrees C, however, the metabolic rates dropped to the same low level, and the core temperature equilibrated with ambient temperature in both genotypes. The results demonstrate that the 1-wk-old fa/fa pup has a defect in thermoregulatory thermogenesis. The magnitude of the deficit in energy expenditure was more than adequate to account for the 50% greater fat content of 7-day-old fa/fa pups.     

Group and individual regulation of physiology and behavior: a behavioral, thermographic, and acoustic study of mouse development

The traditional approach to the study of thermoregulation in young animals focuses on the regulatory capacities of individuals, which, for multiparous species, risks ignoring critical aspects of the early developmental niche. Here, we examined the ontogeny of regulatory behavior in C57BL/6 mice, employing simultaneous behavioral, thermographic, and acoustic measures of groups and individual pups. Litters of mice were placed in a chamber on Postnatal Day (PND) 2, 4, or 8, in which the ambient temperature (T(a)) gradually cycled (over 50 min) from warm (36.5°C) to cool (20°C) and back (to 36.5°C). Litters of all three ages displayed "group regulatory behavior," whereby group size varied with changes in T(a). This coupling, moreover, improved with age. Infrared thermography was used to monitor skin temperature of pups' interscapular (T(IS)) and rump (T(rump)) areas, and to estimate brown adipose tissue (BAT) thermogenesis (T(IS)-T(rump)) in PND4 and PND8 individuals and huddles. Huddling was found to significantly reduce heat loss in pups subject to thermal challenge as groups, compared to pups challenged as individuals. Additionally, females were found to display significantly warmer T(IS) and T(rump) values than male huddlemates. Huddling did not have a consistent effect on emissions of ultrasonic vocalizations, which were generally correlated with ambient temperature and BAT activation. Our results indicate that simultaneous measures of behavioral and physiological response to cooling may prove useful for a variety of applications, including the phenotyping of social dysfunction.     

Huddling by rat pups: ontogeny of individual and group behavior

A full account of behavioral development in rats must include the ontogeny of both individual and group behavior. Most of our accumulated knowledge, however, pertains to individual ontogenesis. Group behavior and its development are readily seen in the huddling behavior of rat pups. A rat huddle is an entity with characteristics and capabilities distinct from those of the individuals that comprise it. The huddle is a natural context for acquiring olfactory preferences for species odors. Olfactory learning in a huddle involves thermal and tactile stimulation from the mother's body but, surprisingly, not the rewards of suckling or of milk transfer. Although there is complete developmental continuity of huddling behavior, the sensory controls of huddling change dramatically during the first 2 weeks of postnatal life. Huddling behavior is initially controlled by thermal cues ("physiological huddling") and then becomes dominated by olfactory stimuli ("filial huddling"). The complex group behavior of huddling was modeled successfully with computational methods. Group behavior emerges from individual interactions, guided entirely by rules of individual behavior (no rules for group behavior). Three simple rules of autonomous activity/inactivity can spawn the patterns of aggregon formation displayed by groups of 7-day-old pups, but not by 10-day-olds. The developmental change evident by Day 10 requires adding a rule by which each individual is affected by the activity state of adjacent pups. Group behavior responded to manipulations of central oxytocin on Day 10, but not on Day 7.     

The effects of litter size during gestation and lactation on rat development prior to weaning

Physiological and behavioral development of rats was affected by prenatal nutrition and postnatal litter size. Prenatal nutrition was manipulated by combining differences in maternal nutrition with variation of prenatal litter size produced by pre-mating isolation of 1 uterine horn. The pups were reared in postnatal litters of either 4 or 12. During the 1st postnatal week, development and free behavior were affected only by the prenatal treatments. During the 2nd postnatal week, both prenatal and postnatal effects were observed. Development and behavior during the 3rd postnatal week reflected only postnatal litter size. Brain weight at weaning was influenced by both the prenatal and postnatal treatments.     

Thermoregulatory competence and behavioral expression in the young of altricial species—Revisited

The behavioral and physiological thermoregulatory capabilities of newborn and infant mammals have been studied for over half a century. Psychobiologists have noted that the infants of altricial species (e.g., rats) have physical and physiological limitations such that heat loss overwhelms heat production, thus forcing a reliance on behavioral thermoregulation for the maintenance of body temperature. Recent evidence, however, suggests that a modification of this view is justified. Specifically, throughout a range of moderately cold air temperatures, nonshivering thermogenesis by brown adipose tissue contributes significantly to the infant rat's behavioral and physiological adaptations to cold challenge. Given the prominent use of altricial species for the study of infant behavior, increased understanding of the infant's physiological responses to cold and the effect of thermal factors on behavior is warranted. © 1998 John Wiley & Sons, Inc. Dev Psychobiol 33: 107–123, 1998     

Unloading does not increase brown-adipose-tissue activity in rat pups

Our previous study has shown that chronic hindlimb suspension leads to an increase in both the thermogenic capacity and the activity in brown adipose tissue (BAT) of adult rats (Yamashita, H., Ohira, Y., Wakatsuki, T., Yamamoto, M., Kizaki, T., Oh-ishi, S., Sato, Y. and Ohno, H. (1995). J. Appl. Physiol. 78: 384-387). In order to examine if unloading also increases the BAT activity in rat pups, the hindlimbs in the suspended pups were unloaded by tail suspension beginning on postnatal day 4 and suspended until day 21. The thermogenic activity (which was assessed by guanosine 5'-diphosphate binding to BAT mitochondria) was markedly lower in 21-day-old suspended pups than in 21-day-old control pups, although there was no difference in uncoupling protein 1 (UCP1) content or UCP1 mRNA expression in the BAT mitochondrial fraction between both pups. Likewise, there was no disparity in either adrenal or thymus mass between the control and suspended pups throughout the experiment. These results suggest that, in contrast to adult rats, chronic hindlimb suspension leads to a decrease in the thermogenic activity in BAT of rat pups possibly for reason that pups are less susceptible to the stress of unloading.     

Ontogeny of behavioral inhibition induced by unfamiliar adult male conspecifics in preweanling rats.

Previous studies showed that when socially isolated at 22 degrees C, postnatal day 14 rats, but not younger day 7 rats, reduce their emission of ultrasonic vocalizations when exposed to an unfamiliar adult male rat, a naturalistic threat. Because ultrasound production is associated with factors such as age and body temperature, this study examined in age-appropriate thermoneutral temperature ranges whether preweanling rats of different ages are equally capable of inhibiting their emission of ultrasounds when threatened. In Experiment 1, 7- and 14-day-old rats were socially isolated and exposed to unfamiliar anesthetized adult male rats in a thermoneutral environment. Only 14-day-old rats significantly reduced their emission of ultrasounds. This reduction in ultrasound production was accompanied by freezing. In Experiment 2, additional ages were examined under identical test conditions. At 3, 6, and 9 days of age, pups frequently emitted ultrasounds when exposed to the anesthetized male rat. However, at 12 days of age, rat pups responded to the anesthetized male rat by freezing and significantly reducing their emission of ultrasounds. Results indicate clearly that under the present testing conditions the ability of rat pups to inhibit ultrasounds and freeze when threatened is not present at birth but emerges by the end of the second postnatal week.     

Activation of brown adipose tissue thermogenesis increases slow wave sleep in rat

Considering the thermoregulatory role of slow wave sleep (SWS), we wondered whether the sole increase of brown adipose tissue (BAT) thermogenesis could enhance this sleep state. We tested this hypothesis by administering to rats an agonist (BRL 37,344) of the beta-3 adrenoceptor subtype that is massively localized in BAT cell membrane and that is known to activate BAT thermogenesis. Sleep was electrographically characterized. The temperature of interscapular BAT (Tibat) and cortex (Tco) were also assessed. Tibat significantly increased 2-3 h after BRL injection (but not Tco), concomitantly with SWS (+56-57%). At the maximum of Tibat, a significant positive correlation was found between their changes and those of SWS. We demonstrated for the first time that sleep (and especially SWS) can be affected by the specific activation of BAT.     

Changes in the categorization of appetitive and aversive events during postnatal development of the rat

Environmental stimuli are not clearly categorized into appetitive and aversive classes during the first postnatal week. Three- and 6-day-old rats are highly activated by nominally appetitive (milk infusions) and aversive (foot shock) stimuli. Both evoked the same generalized behavioral responses (rolling, curling). By 12 days of age, these stimuli were responded to differently; mouthing and probing occurred in response to milk while flinching and escape responses were observed to shock. The affective properties of mild shock were found to be hedonically opposite for 6- and 12-day-old pups. Six-day-old showed an acquired preference for an odor paired with shock which increased with intensity (.1–.5 mA) similar to that observed with milk infusions. An acquired odor aversion was displayed by 12 days old using these shock parameters. These results suggest that young rat pups may not differentiate many appetitive and aversive events behaviorally or affectively until the second postnatal week.     

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.     

Gender markedly modulates behavioral thermoregulation in a non-human primate species, the mouse lemur (Microcebus murinus)

Age and gender are known to significantly modulate thermoregulatory capacities in mammals, suggesting strong impacts on behavioral adjustments, which are used to minimize the energy costs of thermoregulation. We tested the effects of sex and age on spontaneous choice of ambient temperature (Ta) in a non-human primate species, the mouse lemur (Microcebus murinus). The animals acclimated to both winter and summer photoperiods, two seasons significantly modifying thermoregulation function, were experimented in a thermal gradient device.During winter, adult males did not show preference for warm Tas whereas old males did. In contrast, female mouse lemurs of both age categories exhibited great preferences for warm Tas. Acclimation to summer revealed that males selected colder Ta for the day than during the night. Such behavior did not exist in females. Old females explored and selected warmer nests than adult ones.This study raised novel issues on the effect of gender on thermoregulatory capacities in the mouse lemur. Females probably use behavioral adjustments to limit energy expenditure and might prefer to preserve energy for maternal investment by anticipation of and during the breeding season. Further experiments focusing on female thermoregulatory capacities are needed to better understand the energy challenge that may occur during winter and summer in female mouse lemurs, and whether this trade-off changes during aging.     

Development of the circadian rhythm of neuronal activity in suprachiasmatic nucleus of rat hypothalamic slices

Development of the circadian rhythm of neuronal activity in the suprachiasmatic nucleus (SCN) was studied in rat hypothalamic slices. The firing rate of SCN neurons of 7- and 11-day-old rat pups was low during all of the day, whereas that of 14- and 21-day-old pups was high during daytime and low during nighttime. The present results suggest that the circadian rhythm of SCN neuronal activity is established between 11 and 14 days of age, corresponding closely to the time of onset of the other various hormonal and behavioral circadian rhythms.     

Effects of chronic and acute methylphenidate hydrochloride (Ritalin) administration on locomotor activity, ultrasonic vocalizations, and neuromotor development in 3- to 11-day-old CD-1 mouse pups.

The present study examined the effects of chronic and acute treatment with methylphenidate hydrochloride (Ritalin) on isolation-induced ultrasonic vocalizations, spontaneous locomotor activity, and neuromotor coordination in 3- to 11-day-old CD-1 mouse pups. In Experiment 1, 3- to 11-day-old pups received daily injections of saline, 5 mg/kg or 20 mg/kg of methylphenidate hydrochloride, or no injection and were tested on postnatal Days 3, 5, 7, 9, and 11. Both doses of methylphenidate resulted in significant increases in locomotor activity at all ages, but had no significant effect on body weight, neuromotor development, or emission of ultrasonic vocalizations. In Experiment 2, pups were given a single dose of methylphenidate (5 or 20 mg/kg), saline, or no injection on one of postnatal Days 5, 7, 9, or 11. This acute methylphenidate treatment increased locomotor activity, but had no significant effects on ultrasonic vocalizations or neuromotor coordination. These results indicate that short-term, chronic methylphenidate treatment elevates locomotor responses, but has no immediate effects on anxietylike responses or on the development of neuromotor behavior of CD-1 mice in the first 11 days of life.     

Maternally separated rats show deficits in maternal care in adulthood

Although there is considerable research on the phenomenology, neuroendocrinology, neuroanatomy, and sensory control of maternal behavior, little is known about the influences of early postnatal and postweaning experiences on the development of maternal behavior. The purpose of this study was to assess how early life separation from the mother rat affects development of the offspring's juvenile and adult maternal behavior. From postnatal Days 1 to 17, 3 female rats within each litter were separated (SEP) from the mother and the rest of the litter for 5 hr daily while 3 of their sisters were not maternally separated (NSEP). On postnatal Day 21, all subjects were weaned and randomly assigned to one of three juvenile conditions. One female from both SEP and NSEP groups was either isolated (I), given a social conspecific (S), or given 1- to 4-day-old pups (P) for 5 consecutive days. Maternal behavior of SEP and NSEP animals was assessed and recorded on each of the 5 days. Once all animals reached adulthood, they were mated, gave birth, and were assessed for their maternal behavior. We found that the effects of maternal separation on juvenile maternal-like behaviors were minimal. On the other hand, maternal separation reduced adult maternal licking and crouching over pups. In addition, there was a significant interaction between postnatal and juvenile experience on maternal crouching in maternal animals. These results are discussed in terms of the variety of possible behavioral, endocrine, and neurochemical mechanisms that mediate the effects of early life experiences on adult maternal behavior.     

Sensory regulation of maternal behavior in rats: Effects of pup age

Maternal behavior (retrieving, crouching, and licking) was induced in Sprague-Dawley virgin female rats by constant exposure to pups aged 1-2, 3-4, 5-6, 7-8, 9-10, 11-12, 13-14, or 15-16 days. The incidence of spontaneous components of maternal behavior, notably retrieving, was geater towards pups 1-8 days of age than towards older pups, whereas the occurrence of cannibalism did not differ as a function of pup age. With pups 1-2 through 13-14 days, the latency to onset of full maternal behavior was shortest with 1-2-day-old pups (2-day median) and longest with 13-14-day-old pups (7-day median). Females exposed to pups aged 3-4 through 11-12 days did not differ significantly in their latencies, the medians of which ranged from 4.0 to 5.5 days. Only 1 female out of 8 exposed to pups aged 15-16 days became fully maternal, but 5 more displayed components of maternal responsiveness. The optimal nature of neonates and the general attractiveness of a wider range of pup ages as stimuli for the elicitation of maternal behavior in rats, as well as comparisons to mice and hamsters, were discussed.     

Changes in [14C]deoxyglucose incorporation into rat brain regions during heat-seeking behavior in the cold environment

The central nervous structures involved in behavioral thermoregulatory responses during cold exposure were investigated in conscious rats by means of the 2-deoxy-D-[14C]glucose ([14C]-DG) autoradiographic technique. According to autoradiographs, many brain regions with significant increases or decreases in [14C]-DG incorporation were observed during thermoregulatory behavior. When animals were only exposed to cold, significant increases in [14C]-DG incorporation were observed in the caudate putamen, lateral preoptic area, medial forebrain bundle, ventromedial hypothalamus (VMH), posterior hypothalamus, ventroposteromedial thalamus (VPM), dorsomedial thalamus (MD), substantia nigra (SN) and red nucleus (RN). Lower activities were observed in the hippocampus and the medial habenula. However, when animals performed the behavioral thermoregulation (heat seeking behavior) significant increases were noted in the sulcal prefrontal cortex, sensory-motor cortex, and MD, while decreases were noted in the piriform cortex, VMH, VPM, medial habenula, SN and RN, compared with those in the group without the thermoregulatory behavior.     

Increased formalin-induced pain and expression of fos neurons in the lumbar spinal cord of prenatally stressed infant rats

When pregnant dams are stressed, there is a resultant alteration in brain development and behavior in their offspring. Prior work has shown increased nociceptive responses in adolescent or adult rats born of stressed dams. However, the age at which those changes first occur is not known. The aim of the present study was to evaluate the effect of prenatal stress on pain sensitivity in the formalin test in 7-day-old rats, behaviorally and by fos-like immunoreactivity (Fos-LI) in the lumbar spinal cord dorsal horn. The behavioral response to intraplantar injection of formalin is represented by two nociceptive phases separated by an interphase during which nociceptive responses decrease; the interphase is not seen until the start of the third postnatal week and appears as descending inhibitory monoaminergic systems develop. Prenatally stressed infants showed increased nociceptive responses in the second, tonic phase and a large increase in the number of formalin-induced Fos-LI neurons in the lumbar dorsal horn, a result consistent with the behavioral data. The increased nociception in prenatally stressed 7-day-old pups may be associated with the decrease in the intensity of serotonin-like immunoreactivity and density of serotonergic cells in the lumbar spinal cord dorsal horn and the dorsal raphe nucleus reported earlier.