Milk-induced, opioid-mediated antinociception in rats at the time of cesarean delivery.

Four experiments were conducted in rats within 2 hr of cesarean delivery to assess antinociception at birth and its possible opioid bases. Morphine antinociception was established in a dose-dependent fashion (0.0625-5.0 mg/kg bw). Analgesia was naloxone (1.0 mg/kg) reversible. In succeeding experiments, antinociception equivalent to that produced by 0.0625-0.125 mg/kg morphine injections was induced by a single 20-microliters bolus of milk (commercial half-and-half) that was delivered over 1-2 s to the middle of the tongue. This, too, was naloxone reversible. Milk-induced antinociception was maintained for at least 4 min. Finally, baseline latencies were progressively reduced during the first 2 hr after delivery to levels (8-10 s) that are typically obtained in older (10-day-old) rats. This decline was not opioid mediated because it was not affected by naloxone. Thus, at birth, delivering milk to the mouth in physiological volumes can exert opioid-mediated antinociceptive effects in rats born by cesarean delivery that had never suckled or experienced any other form of maternal contact.     

Dopaminergic modulation of local network activity in rat prefrontal cortex

Dopamine modulates prefrontal cortex excitability in complex ways. Dopamine's net effect on local neuronal networks is therefore difficult to predict based on studies on pharmacologically isolated excitatory or inhibitory connections. In the present work, we have studied the effects of dopamine on evoked activity in acute rat brain slices when both excitation and inhibition are intact. Whole cell recordings from layer II/III pyramidal cells under conditions of normal synaptic transmission showed that bath-applied dopamine (30 microM) increased the outward inhibitory component of composite postsynaptic currents, whereas inward excitatory currents were not significantly affected. Optical imaging with the voltage-sensitive dye N-(3-(triethylammonium)propyl)-4-(4-(p-diethylaminophenyl)buta-dienyl)pyridinium dibromide revealed that bath application of dopamine significantly decreased the amplitude, duration, and lateral spread of activity in local cortical networks. This effect of dopamine was observed both with single and train (5 at 20 Hz) stimuli. The effect was mimicked by the D1-like receptor agonistR(+)-6-chloro-7,8-dihydroxy-1-methyl-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine hydrobromide (1 microM) and was blocked by R(+)-7-chloro-8-hydroxy-3-methyl-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine hydrochloride (10 microM), a selective antagonist for D1-like receptors. The D2-like receptor agonist quinpirole (10 microM) had no significant effect on evoked dye signals. Our results suggest that dopamine's effect on inhibition dominates over that on excitation under conditions of normal synaptic transmission. Such neuromodulation by dopamine may be important for maintenance of stability in local neuronal networks in the prefrontal cortex.     

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.     

Differential role of mu, delta and kappa opioid receptors in ethanol-mediated locomotor activation and ethanol intake in preweanling rats

The opioid system modulates ethanol intake and reinforcement in adult and preweanling rodents. While adult heterogeneous rats normally do not show ethanol-mediated locomotor stimulation, preweanling rats show it quite clearly. We recently observed that naloxone, a non-specific opioid antagonist, attenuated ethanol-induced locomotor activation in preweanling rats. In the present study we tested the role of specific opioid receptors (mu, delta and kappa) in ethanol-mediated locomotor stimulation and ethanol intake. In Experiment 1 13-day-old rats received naloxonazine (mu antagonist: 0, 7.5 or 15 mg/kg), naltrindole (delta antagonist: 0, 2 or 4 mg/kg) or nor-binaltorphimine (kappa antagonist: 0, 2, 4 or 8 mg/kg) before an intragastric administration of ethanol (0 or 2.5 g/kg), and subsequent locomotor activity assessment. In Experiment 2, the same opioid antagonists were administered on postnatal days 13 and 14 before consumption of ethanol (6%), saccharin (0.05%) or distilled water. In Experiment 1 only naloxonazine reduced ethanol-mediated locomotor stimulation. None of the opioid antagonists affected locomotor activity in water controls. In Experiment 2 naloxonazine and naltrindole suppressed ingestion of all the solutions tested. Similar to what has been reported in adult rodents, mu-opioid receptors seem to modulate ethanol-activating effects during early ontogeny. Hence, there seems to be a partial overlap of neurochemical mechanisms involved in the rewarding and stimulating effects of ethanol in preweanling rats. Mu-receptor antagonists reduced both ethanol-induced activity and ethanol intake, but it is unclear whether the latter effect is specific to ethanol or only a reflection of an effect on consummatory behavior generally, since mu and delta receptor antagonists also suppressed ingestion of water and sacccharin.     

Locomotor activity, ultrasonic vocalization and oxytocin levels in infant CD38 knockout mice

Oxytocin (OT), a neurohormone involved in reproduction, plays a critical role in social behavior in a wide range of mammalian species from rodents to humans. The role of CD38 in regulating OT secretion for social behavior has been demonstrated in adult mice, but has not been examined in pups or during development. Separation from the dam induces stress in 7-day-old mouse pups. During such isolation, locomotor activity was higher in CD38 knockout (CD38(-/-)) pups than in wild-type (CD38(+/+)) or heterozygous (CD38(+/-)) controls. The number of ultrasonic vocalizations was lower in CD38(-/-) pups than in CD38(+/+) pups. However, the difference between the two genotypes was less severe than that in OT knockout or OT receptor knockout mice. To explain this, we measured plasma OT levels. The level was not lower in CD38(-/-) pups during the period 1-3 weeks after birth, but was significantly reduced after weaning (>3 weeks). ADP-ribosyl cyclase activities in the hypothalamus and pituitary were markedly lower from 1 week after birth in CD38(-/-) mice and were consistently lower thereafter to the adult stage (2 months old). These results showed that the reduced severity of behavioral abnormalities in CD38(-/-) pups was due to partial compensation by the high level of plasma OT.     

Dopaminergic activity modulation via aggression, status, and a visual social signal

Social interaction may elicit aggression, establish social rank, and be influenced by changes in central dopaminergic activity. In the lizard Anolis carolinensis, a sign stimulus (darkening of postorbital skin or eyespots) inhibits aggressive response from opponents, in part because it forms more rapidly in dominant males. The authors report that artificially hiding or darkening eyespots influences central dopaminergic activity, social status, and aggression during dyadic social interaction. All males that viewed an opponent with eyespots painted black became subordinate and exhibited elevated dopamine in raphe, lateral amygdala, and medial amygdala but decreased dopamine in septum and locus ceruleus. In contrast, males that viewed opponents with hidden eyespots (painted green) became dominant and had increased dopamine in striatum, nucleus accumbens, hypothalamus, and combined substantia nigra/ventral tegmental area.     

Olfactory, thermal, and tactual influences on infantile ultrasonic vocalization in rats.

The amount of time infant rats 3-13 days old spent emitting ultrasounds was greater when pups were placed in an empty dish than when placed in a dish containing clean bedding or soiled bedding from the nest. Pups from 5-13 days of age vocalized more when on the clean bedding than when on the soiled bedding. When placed on cloth covered dishes, pups vocalized most to the empty dish and least to the dish containing soiled bedding. The odor of clean bedding elicited less ultrasonic vocalization than the odor of the empty dish only when less than 7 days old. Pups vocalized more to room temperature than to heated conditions. The results indicate that olfactory, tactual, and thermal nest cues influence infantile ultrasonic vocalization in the rat.     

Within-species variation in the development of ultrasonic signaling of preweanling rats.

The development of litter and individual differences in the rate of ultrasonic signaling of neonatal rats was studied. Systematic variations among litters and individuals emerged, without differential treatment. These differences were not correlated with variations in general development as indexed by body weight. Two experiments using a cross-fostering design showed that litter differences developed independently of variations in postnatal environment. These results indicate that the variations among litters in ultrasound rate have a prenatal, possibly genetic, etiology and may represent reliable indicants of response to environmental stress.     

Dopaminergic modulation of nNOS expression in the pituitary gland of male rat

Nitric oxide is an unconventional transmitter since it is not transported and released by exocytosis. In the pituitary gland, nitric oxide is locally synthesised by gonadotroph and folliculo-stellate cells. Dopamine, the principal central inhibitory signal in prolactin release, may exert its inhibitory effects by stimulation of nitric oxide production. However, the effects of dopaminergic modulation on nitric oxide-producing pituitary cells have not been analysed. Therefore, we examined the effects of intraventricular administration of the dopamine antagonist haloperidol (40 g) on the pituitary expression of neuronal nitric oxide synthase (nNOS) in male adult rats. In untreated and control animals, nNOS-positive cells were very similar. Two types of nNOS-positive cells appeared in the pars distalis: round or polygonal cells and stellate cells. Although some isolated cells were found, the nNOS-positive cells commonly appeared grouped in clusters close to blood vessels. nNOS immunoreactivity appeared as a uniform staining throughout the cytoplasm, including cell prolongations. The number and size of nNOS-expressing cells in the pituitary gland decreased significantly after treatment with haloperidol (p<0.01). To evaluate the potential direct effect of dopamine on pituitary cells, pituitary monolayer cultures were treated with dopamine during a time-course of 12 h. Our in vitro studies revealed that dopamine increases the percentage of nNOS-positive cells and augments cellular area (p<0.05). These results demonstrate that: (1) treatment of rats in vivo with a dopamine antagonist significantly decreases expression of nNOS in the pituitary and (2) in vitro dopamine exerts a direct effect on pituitary cultures by increasing nNOS-positive cells. Thus, these findings suggest that dopamine may function as a physiological stimulator of nNOS expression in the rat pituitary gland.Sources of support in the form of grants: Junta de Castilla y León (SA58/98, SA05/99, SA50/99), DGES (PB97-1341; HI1998-0160), FIS (99/1187), CICYT (1FD97-1325) and BIOMED I program of the EU (BMH1-CT94-1536)     

Role of dopamine D(1) receptors for kappa-opioid-mediated locomotor activity and antinociception during the preweanling period: a study using D(1) receptor knockout mice

kappa-Opioid receptor agonists both increase the locomotor activity of preweanling rats and induce antinociception. To determine whether dopamine (DA) D(1) receptors are necessary for either of these kappa-opioid-mediated effects we used D(1) (D(1A)) receptor knockout mice (i.e., D(1)-deficient mice). Heterozygous, wild-type, and D(1)-deficient mice (13 days old at testing) were injected with the kappa-opioid receptor agonist U-50,488 methanesulfonate (0.0, 0.2, 1. 0, 2.5, or 5.0 mg/kg, s.c.) and locomotor activity was measured for 60 min. In a separate experiment, tail-flick latencies of heterozygous, wild-type, and D(1)-deficient 13-day-old mice were assessed both before and after treatment with U-50,488 (0.0, 1.0, 2. 5, 5.0, or 10.0 mg/kg, s.c.). Results showed that lower doses of U-50,488 (0.2 and 1.0 mg/kg) increased the locomotor activity of 13-day-old mice regardless of genotype. Besides affecting locomotion, kappa-opioid receptor stimulation induced antinociception in preweanling mice, as U-50,488 caused a dose-dependent increase in the tail-flick latencies of heterozygous, wild-type, and D(1)-deficient mice. U-50,488's locomotor activating and analgesic effects did not differ according to genotype, thus suggesting that D(1) receptors are not necessary for kappa-opioid-mediated locomotor activity and antinociception during the preweanling period.     

Influence of neonatal handling on blood pressure,locomotor activity,and preweanling heart rate in spontaneously hypertensive and Wistar Kyoto rats

This experiment tested the hypothesis that increased stimulation early in development would (a) alter developmental changes in heart rate and behavioral reactivity and (b) affect the level at which blood pressure was regulated in adulthood. For this purpose, the effects of daily handling and maternal separation (3 min per day) on both behavioral and cardiovascular measures were examined in spontaneously hypertensive (SHR) and normotensive control Wistar Kyoto (WKY) rats. Prior to weaning, elevated heart rates in pups handled during the first postnatal week were most pronounced among 4-week-old prehypertensive SHR pups. Early handling affected behavior observed during openfield testing similarly in young adult rats of the SHR and WKY strains (e.g., increased locomotor activity on the first day of testing). In female rats of the WKY strain, early handling resulted in a lower baseline blood pressure; the blood pressure; the blood pressure of SHR rats was not affected by increased stimulation in infancy. Examination of longitudinal data yielded no support for a direct association between behavioral reactivity or preweaning heart rate and high blood pressure. These findings demonstrate the influence of both early environmental conditions and genetic factors on maturation within the cardiovascular system and suggest that genetic models of pathological conditions may provide a productive means of examining environmentally shaped aspects of individual differences in physiological regulation.     

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.     

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     

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.     

N-methyl-D-aspartate and dopamine receptor involvement in the modulation of locomotor activity and memory processes

In this study we report on the effects of N-methyl-d-aspartate (NMDA)- and dopamine (DA)-receptor manipulation on the modulation of one-trial inhibitory avoidance response and the encoding of spatial information, as assessed with a non-associative task. Further, a comparison with the well-known effects of the manipulation of these two receptor systems on locomotor activity is outlined. It is well assessed that NMDA-receptor blockage induces a stimulatory action on locomotor activity similar to that exerted by DA agonists. There is evidence showing that the nucleus accumbens is involved in the response induced by both NMDA antagonists and DA agonists. We show results indicating a functional interaction between these two neural systems in modulating locomotor activity, with D2 DA-receptor antagonists (sulpiride and haloperidol) being more effective than the D1 antagonist (SCH 23390) in blocking MK-801-induced locomotion. A different profile is shown in the effects of NMDA antagonists and DA agonists in the modulation of memory processes. In one-trial inhibitory avoidance response, NMDA antagonists (MK-801 and CPP) impair the response on test day, while DA agonists exert a facilitatory effect; furthermore, sub-effective doses of both D1 (SKF 23390) and D2 (quinpirole) are able to attenuate the impairing effect in a way similar to that induced by NMDA antagonists. The effects of NMDA- and DA-acting drugs on the response to spatial novelty, as assessed with a task designed to study the ability of animals to react to discrete spatial changes, are in good accord with the effects observed on passive avoidance. The results show that NMDA as well as DA antagonists, at low doses, selectively impair the reactivity of mice to spatial changes. In a last series of experiments, the possible role of NMDA receptors located in the nucleus accumbens was investigated regarding reactivity to spatial novelty. The experiments gave apparently contrasting results: while showing an impairing effect of focal administrations of NMDA antagonists in the nucleus accumbens on reactivity to spatial novelty, no effect of ibotenic acid lesions of the same structure was observed.     

Thermogenesis during ultrasonic vocalization by rat pups isolated in a warm environment: a thermographic analysis.

Ultrasonic vocalizations, emitted by rat pups when separated from their mother, littermates, and home cage, have been used as a measure of isolation distress. Recently, we demonstrated that cold exposure is the primary component of isolation that induces the vocalization. We were unable, however, to suppress all ultrasound production when transferring pups to a thermoneutral (35 degrees C) environment. Using an infrared thermography system that allows us to estimate noninvasively heat production by brown adipose tissue, we found that pups transferred from the home nest to a 35 degree C test chamber exhibited sizable levels of heat production while they were vocalizing. Moreover, both heat production and ultrasound emission decreased over the 15-min test. Next, we used extreme care to minimize thermal, and therefore respiratory, stimulation of pups before, during, and after the transfer procedure. We found that such precautions prevented both heat production and ultrasound emission following transfer. These results indicate that infant rats' thermal sensitivities are far greater than previously suspected.     

Acquisition and retention of habituation in the preweanling rat

The acquisition and retention of habituation to air-puff was studied in rats on Days 1-19 postpartum. Animals of all ages demonstrated habituation but the rate of habituation varied with age: slower on each successive day from Day 1 to Day 8, but thereafter more rapid on each successive day. This peak of elicited responding is discussed in relation to the peak of spontaneous activity at Day 16 and to the pattern of neural development, including changes in acetylcholinesterase, norepinephrine, and serotonin, measured during the preweaning period. All ages showed 30-min retention of habituation, with 60-min retention developing after the 1st week. No evidence of 24-hr retention appeared at any age.     

Subglottal pressure, tracheal airflow, and intrinsic laryngeal muscle activity during rat ultrasound vocalization

Vocal production requires complex planning and coordination of respiratory, laryngeal, and vocal tract movements, which are incompletely understood in most mammals. Rats produce a variety of whistles in the ultrasonic range that are of communicative relevance and of importance as a model system, but the sources of acoustic variability were mostly unknown. The goal was to identify sources of fundamental frequency variability. Subglottal pressure, tracheal airflow, and electromyographic (EMG) data from two intrinsic laryngeal muscles were measured during 22-kHz and 50-kHz call production in awake, spontaneously behaving adult male rats. During ultrasound vocalization, subglottal pressure ranged between 0.8 and 1.9 kPa. Pressure differences between call types were not significant. The relation between fundamental frequency and subglottal pressure within call types was inconsistent. Experimental manipulations of subglottal pressure had only small effects on fundamental frequency. Tracheal airflow patterns were also inconsistently associated with frequency. Pressure and flow seem to play a small role in regulation of fundamental frequency. Muscle activity, however, is precisely regulated and very sensitive to alterations, presumably because of effects on resonance properties in the vocal tract. EMG activity of cricothyroid and thyroarytenoid muscle was tonic in calls with slow or no fundamental frequency modulations, like 22-kHz and flat 50-kHz calls. Both muscles showed brief high-amplitude, alternating bursts at rates up to 150 Hz during production of frequency-modulated 50-kHz calls. A differentiated and fine regulation of intrinsic laryngeal muscles is critical for normal ultrasound vocalization. Many features of the laryngeal muscle activation pattern during ultrasound vocalization in rats are shared with other mammals.     

Dissociation of systemic and hippocampal modulation of rat locomotor activity by 5-HT(2C) receptors

In the present study, the ability of 5-hydroxytryptamine(2C) (5-HT(2C)) receptors in the hippocampus to enhance locomotor activity in rats was investigated by local infusion. Intraperitoneal injection of the selective 5-HT(2C) antagonist SB 242084 (1 mg/kg) significantly increased rats motor activity, while the effects of non-selective 5-HT(2C) agonist m-chlorophenylpiperazine (m-CPP, 10 mg/kg; i.p.) on motor activity were lower than those of the control group. In the day hours, the local infusion of non-selective 5-HT(2C) agonist, m-CPP (1.0 mM) into the bilateral hippocampus via microdialysis probes increased locomotor activity in contrast with intraperitoneal injection. This increase was completely reversed by the combined infusion of the selective 5-HT(2C) antagonist SB 242084 (100 mM). In the night hours, the local infusion of SB 242084 (100 mM) into the bilateral hippocampus significantly inhibited the nocturnal hyperactivity, which was reversed by the combined infusion of m-CPP. The present study demonstrates that the 5-HT(2C) receptors in the hippocampus act to increase rat locomotor activity.     

Circadian rhythms of locomotor activity in the subterranean Mashona mole rat, Cryptomys darlingi

The Mashona mole rat, Cryptomys darlingi, is a social, subterranean African rodent that is rarely, if ever, exposed to light, and that exhibits a regressed visual system. This study investigated locomotor activity patterns of Mashona mole rats (n=12) under different light cycles. Activity was measured using either infrared captors (n=8) or running wheels (n=4). The mole rats entrained their activity to a standard (LD 12:12) photoperiod. They displayed either a nocturnal or diurnal activity preference with one bout of activity and one bout of rest. Therefore, as a species, the Mashona mole rat did not show a clear nocturnal or diurnal activity preference. When the LD (12:12) light cycle was inversed, the animals switched their activity, too. Under constant dark (DD), most mole rats (73%) showed a free-running circadian activity rhythm, but under constant light (LL), only some (36%) did. The free-run period of the rhythm (tau) ranged from 23.83 to 24.10 h. The remaining animals were arrhythmic. There was large interindividual and intraindividual variations in the rate and extent of entrainment, time of activity preference, and activity patterns. Possible reasons for the observed variations are discussed. It is concluded that the Mashona mole rat has an endogenous activity rhythm which approximates 24 h, that the mole rat can distinguish between light and dark, and that the endogenous clock utilises this photic information as a zeitgeber.