Family Nurture Intervention in preterm infants alters frontal cortical functional connectivity assessed by EEG coherence.

AIM: To assess the impact of Family Nurture Intervention (FNI) on cortical function in preterm infants at term age. METHODS: Family Nurture Intervention is a NICU-based intervention designed to establish emotional connection between mothers and preterm infants. Infants born at 26-34 weeks postmenstrual age (PMA) were divided into two groups, standard care (SC, N = 49) and FNI (FNI, N = 56). Infants had EEG recordings of ~one hour duration with 124 lead nets between 37 and 44 weeks PMA. Coherence was measured between all pairs of electrodes in ten frequency bands. Data were summarised both within and between 12 regions during two sleep states (active, quiet). RESULTS: Coherence levels were negatively correlated with PMA age in both groups. As compared to SC infants, FNI infants showed significantly lower levels of EEG coherence (1-18 Hz) largely within and between frontal regions. CONCLUSION: Coherence in FNI infants was decreased in regions where we previously found robust increases in EEG power. As coherence decreases with age, results suggest that FNI may accelerate brain maturation particularly in frontal brain regions, which have been shown in research by others to be involved in regulation of attention, cognition and emotion regulation; domains deficient in preterm infants.

Physiological responses of infant rats to separation from their mothers.

Decreases of 40 percent in cardiac and respiratory rates occur during the first 12 to 16 hours after 2-week-old rat pups are separated from their mothers. These rates decrease without significant alteration in activity level and despite maintenance of body temperature, of nutrition by intubation, of an intact litter, and of the home cage nest.

Cardiac rate regulated by nutritional factor in young rats.

Milk fed by stomach tube to 2-week-old rats separated from their mothers without feeding for 16 hours transiently but fully reversed the decrease in cardiac rate which had occurred during separation. This effect was rapid in onset and was related to dosage; it was not dependent upon gastric distention, but did depend upon beta-adrenergic transmission.

Early maternal separation increases gastric ulcer risk in rats by producing a latent thermoregulatory disturbance.

Rat pups that are separated early from their mothers, at postnatal day 15, become hypothermic when subjected to physical restraint on postnatal day 30. Restraint of separated pups also elicits an unusually high incidence of gastric erosions, as well as insomnia and an increase in quiet wakefulness. If hypothermia during restraint is prevented, neither the erosions nor the behavioral responses occur. Rat pups separated at the customary age (postnatal day 22) do not become hypothermic during restraint, and the restraint of such pups is not associated with either gastric erosion or insomnia.

Vertical transmission of acquired ulcer susceptibility in the rat.

Premature separation of rat pups from their dams greatly increases their susceptibility to restraint-induced gastric erosions. When prematurely separated female rats grow to adulthood and mate with stock males, their normally reared F 1 progeny also have increased susceptibility to restraint-induced erosions. Cross-fostering studies show that prenatal rather than postnatal factors transmit this susceptibility to the F 1 progeny.

Sleep-wake state organization in infant rats with episodic respiratory disturbance following sinoaortic denervation.

The role of arterial chemoreceptors in the maintenance of rhythmic respiration during sleep was investigated in 2-week-old rats by microsurgical sectioning of carotid sinus and aortic depressor nerve fibers (SAD) and later recording of respiratory pattern by impedance pneumography and sleep-wake states by electrophysiological methods. Deafferentation produced high rates of an atypical respiratory pattern characterized by episodes during which the usual pattern was replaced by an arrhythmic series of low amplitude waveforms with frequent short pauses and occasional gasps. These episodes lasted a mean of 20s, occurred 15.1 times/h and were virtually confined to the REM state. Apneas occurred in all stages of sleep and were not increased by SAD but were significantly associated with state changes only after denervation. Body movements were unchanged and sleep-wake state organization was only minimally affected by SAD. These results suggest that the REM state may constitute a vulnerable period for respiratory control in infancy during which afferent feedback from peripheral arterial chemoreceptors may ordinarily play a stabilizing role.

Multiple regulators of ultrasonic vocalization in the infant rat.

This paper reviews our growing understanding of the external environmental and the internal neuroregulatory mechanisms for the infant rat's vocal response to abrupt isolation from nest, littermates and mother. Interactions of the rat pup with specific components of its animate and inanimate environment have been identified and found to exert a tonic regulatory action on the neural systems mediating ultrasonic vocalization (USV). In isolation, withdrawal of these regulators produces an acute outburst of calling. Changes in rate of ultrasonic calling in response to administration of receptor agonist and antagonist agents generally correspond to the effects of those agents on measures of anxiety in adult animals and in human clinical populations, suggesting a conservation of neural substrates for anxiety in evolution and in development. The inhibition of isolation calling in response to predator cues and its potentiation by brief exposure to maternal cues, represent novel forms of regulation with implications for neuroendocrine development and for the neurobiology of early cognitive-emotional processes.

Socially mediated reduction of isolation distress in rat pups is blocked by naltrexone but not by Ro 15-1788.

The presence of a single anesthetized littermate significantly reduced the rate of ultrasonic vocalization by 10-day-old pups isolated in a novel environment. Naltrexone (1.0 mg/kg) returned the vocalization rate to the level of pups tested alone and disrupted the maintenance of body contact between the test pup and a companion. This suggests that the companion exerts comforting effects through endogenous opioid mechanisms. Although chlordiazepoxide is as effective as morphine in the quieting of isolation distress, the benzodiazepine (BDZ) antagonist Ro 15-1788 (5, 10, or 20 mg/kg) was ineffective in blocking the comfort effect and facilitated quiet contact with the companion. In isolated pups, Ro 15-1788 caused a significant, but not a dose-related, decrease in vocalization, a possible indication of the displacement of an endogenous anxiogenic ligand at the BDZ receptor complex.

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.

Trigeminal and olfactory pathways mediating isolation distress and companion comfort responses in rat pups.

Isolation in a novel test cage elicits ultrasonic vocalization (USV) in 12-13-day-old rat pups, and the presence of an anesthetized littermate companion reduces USV while eliciting body contact. These responses were studied in pups with surgical lesions of trigeminal snout afferents and after olfactory denervation by nasal ZnSO4 perfusion. With lesions of only one system, body contact with the passive companion was reduced by 20%, and interpup orientation was differentially affected by the two types of lesions. However, USV rates were no different from littermate controls in the presence of the companion or when tested alone. Combined lesions of both systems reduced contact time 55% and eliminated any reduction in USV rates by the companion while leaving intact USV and locomotor responses to isolation. These results suggest that different sensory pathways may mediate isolation distress and companion comfort responses.

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.

Ultrasonic vocalization, laryngeal braking, and thermogenesis in rat pups: a reappraisal.

In these experiments we tested the hypothesis that the respiratory events underlying rat pup ultrasonic vocalization (USV) facilitate brown adipose tissue (BAT) thermogenesis, as proposed by Blumberg and Alberts (1990). Laryngeal denervation or tracheostomy were used to reduce or eliminate laryngeal braking and prevent USV in 9- to 10-day-old pups during recovery from deep hypothermia and during cooling induced by isolation for 1 hr at room temperature. Both forms of devocalization slowed rewarming of hypothermic pups, but neither had any effects on cooling rate or BAT thermogenesis in normothermic pups that were allowed to cool from 35 degrees C to 29 degrees C. Physiological effects of laryngeal braking on thermogenesis appear to be limited to conditions, such as severe hypothermia, in which ventilatory and oxygen transport functions are compromised. The results have implications for understanding the regulation and evolution of USV.

Independence of benzodiazepine and opiate action in the suppression of isolation distress in rat pups.

To determine whether benzodiazepines (BDZs) quiet isolation distress in 10-day-old rat pups by causing a release of endogenous opioids, a blockade of the effects of chlordiazepoxide (CDP) by the opiate antagonist naltrexone (NLX) was sought. Nonsedating doses of morphine (MOR) (0.125 mg/kg) and CDP (2.0 mg/kg) were equally effective in reducing ultrasonic vocalizations and other indices of isolation distress. Appropriate blocking agents NLX, (0.5 mg/kg) against MOR and Ro 15-1788 (4.0 mg/kg) against CDP returned distress measures to levels of saline-treated rat pups. However, NLX failed to reverse the quieting effects of CDP. If CDP potentiates endogenous opioid release, then NLX should block the CDP effect. A higher dose of CDP did not reveal a release of endogenous opioids, and a higher dose of NLX did not antagonize CDP. The quieting effects of BDZs on isolation distress do not appear to be mediated by the opiate system.

The isolation and companion comfort responses of 7- and 3-day-old rat pups are modulated by drugs active at the opioid receptor.

Rat pups emit ultrasonic vocalizations (USVs) when isolated in a novel environment. In 10-day-olds, USV has been shown to be reduced by either the administration of 0.125 mg/kg morphine (MOR) or the presence of a littermate; these effects were both reversed by naltrexone (NLX), an opioid receptor blocker. The present study reports that the same dose of MOR produced NLX-antagonized quieting without sedation in 7- and 3-day-old pups; higher doses of MOR decreased USV but produced motor deficits as well. The 0.125 mg/kg dose of MOR is less effective in reducing USV in 3- and 7-day-olds; calling rates declined by no more than 42%, compared with 65% at 10 days of age. The presence of a companion also lowered the USV of 3- and 7-day-olds by a lesser amount (55-57%) than the 67% seen in 10-day-olds or the 90% decline when pups are 2 weeks old. This suggests that age-related changes in the opioid system may be relevant to the increased salience of a social companion that comes with maturity.

Maternal interactions prior to separation potentiate isolation-induced calling in rat pups.

The vocal response rates of 12-13-day-old infant rats to isolation in a bare test box are markedly increased by brief (1-min) periods of contact with an anesthetized dam prior to isolation, without affecting other isolation-induced behaviors. No such potentiation followed brief contact with littermates, novel test conditions, or experimenter handling. Brief contact with the dam was equally effective in the test chamber or home cage and was not further enhanced by repeated contact-separation sequences. Passive contact became ineffective when prolonged to 30 min, and potentiation could not be restored by providing the additional reinforcing events of continuous suckling, periodic oxytocin-induced milk letdown, or bouts of simulated maternal licking. However, when pups engaged in active interaction with an awake dam, potentiation was significantly enhanced following 1-, 10-, and 30-min periods. A working hypothesis is outlined for the adaptive role of potentiation in the development of the rat pup.

Ultrasonic vocalizations are elicited from rat pups in the home cage by pentylenetetrazol and U50,488, but not naltrexone.

Although isolated rat pups emit ultrasonic vocalizations (USVs), those kept warm and undisturbed in the home cage with their littermates seldom do. Drugs were administered to 10-day-old pups in the home cage to determine whether pharmacological agents can elicit USV in this familiar environment. Ten-day-old Wistar rats were injected with U50,488, a highly selective kappa opioid agonist; pentylenetetrazol (PTZ), an anxiogenic drug that binds at the GABA-benzodiazepine receptor complex; or naltrexone (NLX), an opiate receptor blocker, and then were returned to their littermates in the home cage. U50,488 increased USV and activity levels, lowered body temperature, and disrupted contact with littermates. PTZ raised activity levels but had a smaller effect on vocalization rates and did not alter temperature or contact with littermates. Behavioral measures and body temperature were unchanged by NLX.

Behavioral mechanisms for active maternal potentiation of isolation calling in rat pups.

The ultrasonic vocalization (USV) response of the isolated infant rat is a promising model for studying the neurobiology of an early anxiety state, and potentiation of the USV response after brief maternal encounters is a newly discovered behavioral regulator of this state. Using experimental variations in the contexts and patterns of maternal behavior during pup encounters and manual simulations of maternal behavior, we have identified several specific mother-pup interactions that cause potentiation. When one of these, pup carrying, was manually simulated, potentiation followed only if a characteristic postural transport response had been fully expressed by the pup. These behavioral mechanisms and other data suggest hypotheses for the development and adaptive role of maternal potentiation.

Differential effects of specific opioid receptor agonists on rat pup isolation calls.

The possibility was investigated that specific opioid receptor types might selectively alter the production of isolation-induced ultrasonic vocalizations. Intracisternal injections of mu, delta and kappa opioid receptor agonists were administered to isolated 10-day-old rat pups. The mu receptor agonist [D-Ala2-NMe-Phe4-Gly-ol]-enkephalin (DAMGO) and delta receptor agonist [D-Pen2, D-Pen5]-enkephalin (DPDPE) both reduced the rate of isolation-induced ultrasonic calling in the absence of sedation. The kappa receptor agonist U50,488 had the opposite effect, significantly raising the rate of vocalization. Fourteen-day-old pups, with a larger delta receptor population, showed a greater sensitivity to DPDPE than was seen in the younger animals.

Lethal respiratory disturbance in neonatal rats after arterial chemoreceptor denervation.

The role of afferent feedback from arterial chemoreceptors in the maintenance of rhythmic respiration during early development was studied by section of carotid sinus and aortic nerves of rat pups at different ages from 3 days to 3 weeks postnatally. This deafferentation produced a severe, episodic respiratory disturbance, limited to pups younger than 21 days and associated with mortality rates near 50% during the 2 weeks following surgery. These findings may have implications for the role of peripheral chemoreceptors in the periodic apneas of premature infants and in the Sudden Infant Death Syndrome.

Nutrient control of cardiac rate in the infant rat: alpha-adrenergic mechanisms.

Changes in level of nutrient intake have been shown to exert a major influence on beta-adrenergic cardiac activity in 2 week old suckling rats. A series of experiments demonstrates that the alpha-adrenergic vasoconstrictor system is functional by 2 weeks postnatal age, and that alpha-adrenergic blockade with phenoxybenzomine (PBZ) does not affect the high heart rates of well-fed pups but prevents development of the bradycardia after cessation of suckling and fully reverses the low heart rates of nutrient-deprived pups in a dose-dependent manner. This last effect is dependent upon beta-adrenergic reflex cardiac pathways. After PBZ, milk feeding no longer produces cardiac acceleration in nutrient-deprived pups. Systolic and mean arterial pressure during PBZ administration and estimates of plasma volume change during nutrient deprivation are consistent with the inference that changes in peripheral resistance, mediated by the alpha-adrenergic vasoconstrictor system, accompany nutrient regulation of cardiac rate in the suckling rat.