Lactogenic hormone regulation of maternal behavior.

Biological factors can profoundly affect a mother's response to her young. For example, it is well known that the hormones of pregnancy act on the maternal brain to stimulate the spontaneous onset of maternal behavior at parturition. Studies in the rat have provided an excellent model to investigate maternal behavior in mammals, since maternal behavior in rats is easily observable and readily quantifiable and it is well-documented that the endocrine state of gestation helps to bring about the onset of maternal behavior around the time of birth. The same response in virgin animals requires a number of days of constant exposure to pups before maternal-like behaviors emerge. To date, research has established that the steroid hormones, estradiol and progesterone, and the lactogenic hormones, prolactin and the placental lactogens, act in concert to stimulate maternal behavior in the pregnant female. Treatment of adult, virgin rats with these hormones can stimulate a rapid onset of maternal care. In the present chapter experiments are described that demonstrate key roles for prolactin and placental lactogens in the onset of maternal behavior. Central sites of action of prolactin and placental lactogens, including the medial preoptic area, appear to be involved in stimulating the onset of maternal care. Other studies are discussed which support the involvement of the prolactin receptor in the endocrine regulation of maternal behavior using prolactin receptor antagonist and 'knock-out' models in rats and mice, respectively. Overall, these studies indicate that during pregnancy the endocrine system primes the mother's brain so that the new mother displays appropriate and successful behaviors toward her newborn at parturition.

Expression of mu-opioid receptor mRNA in the medial preoptic area of juvenile rats.

Previous studies indicate that the latency to initiate parental behavior in both male and female rats increases with age; weanling (21 days old) rats display parental behavior 0-2 days after exposure to newborn pups, while older juveniles (30 days old) require 5-6 days of pup exposure before they express the behavior. Furthermore, activation of mu-opioid receptors inhibits parental behavior in juvenile and adult rats. We hypothesized that the age-related increase in behavioral latency could be modulated by the induction of mu-receptor expression in the medial preoptic area (MPOA), a region in which mu-receptors regulate parental behavior. In situ hybridization histochemistry was used to measure mu-receptor mRNA expression in the MPOA of male and female Sprague-Dawley rats that were 21, 24, 27, 30, or 33 days old. Using autoradiographic film analysis, we observed that neurons within part of the MPOA expressed very dense mu-receptor mRNA. Comparison of mRNA distribution with histological boundaries indicated that neurons within the medial preoptic nucleus (MPN), excluding the central part, exhibited the highest density of mu-receptor mRNA within the MPOA. High densities of mu-receptor mRNA extended dorsolaterally and caudally from the MPN toward the bed nucleus of the stria terminalis. MPN mu-receptor mRNA expression was not altered with age and no sex difference was observed. The dense presence of mu-receptor mRNA in the MPN suggests that ample substrate exists on which mu-receptor ligands could modulate the latency to begin parental behavior in juvenile rats, but such behavioral expression apparently is not mediated by a change in mu-receptor mRNA production.

The development of POMC gene expression in the medial basal hypothalamus of prepubertal rats.

Alterations in brain opioid gene expression may underlie the dramatic change in the latency to display parental behavior in juvenile rats. Male and female juvenile rats (18-25 days of age) exhibit parental behavior either immediately or within 1-2 days after coming in contact with foster pups. By 30 days of age, however, their response latencies increase to adult levels of 5-10 days. Given the established involvement of the endogenous opioid system in adult maternal and juvenile parental behaviors, the objective of the present report was to determine possible changes in proopiomelanocortin (POMC) gene expression in the medial basal hypothalamus (MBH) during this early developmental window. We compared POMC gene expression in the MBH of male and female juvenile rats from 21 to 33 days of age by in situ hybridization histochemistry. A significant increase in the number of POMC cells in males and females was detected at 30 days of age in the central portion of the arcuate nucleus. This increase in POMC mRNA may contribute to the shift in parental behavior that occurs in male and female juvenile rats.

Hypothalamic involvement in the regulation of maternal behaviour in the rat: inhibitory roles for the ventromedial hypothalamus and the dorsal/anterior hypothalamic areas.

The present report examines the possible involvement of the ventromedial hypothalamus (VMH), the dorsal hypothalamus (DH), and the anterior hypothalamic area (AHA) in the regulation of maternal behaviour in the female rat. In a series of experiments it was found that either infusions of saline or lowering cannulas into the VMH stimulated a rapid onset of maternal behaviour in progesterone plus oestrogen-primed, nulliparous rats. The stimulatory effect of cannula lowering into the VMH on maternal behaviour was shown to be steroid-dependent. Next, the involvement of cell bodies located in the DH/AHA in maternal behaviour was examined after bilateral lesions of these regions with the neurotoxin, N-methyl-D-aspartic acid (NMA). NMA lesions of the DH/AHA stimulated a rapid onset of maternal behaviour in oestrogen-treated, nulliparous rats, while NMA lesions in non-steroid-treated animals or vehicle infusions in steroid or non-steroid-treated rats failed to induce a rapid onset of behaviour. In a final study the effects of NMA lesions of the VMH were evaluated. As in AHA lesioned rats, NMA lesions of the VMH stimulated a fast onset of maternal behaviour in steroid-primed females. These findings indicate that the VMH as well as the DH/AHA exert chronic steroid-dependent inhibitory influences on the induction of maternal behaviour.

Reproductive experience and opioid regulation of luteinizing hormone release in female rats.

The objective of the present study was to determine whether reproductive experience that produces shifts in opioid regulation of prolactin secretion and behavioural functions also alters opioid regulation of LH during the oestrous cycle or lactation. In Expt 1 the effect of naloxone administration (i.v.) on LH was compared between age-matched, nulliparous and primiparous, catheterized female rats on dioestrus II. In Expt 2, the effects of multiple reproductive experiences on opiate control of LH were investigated using cyclic, nulliparous and multiparous (three litters) rats. In both experiments, no differences in naloxone-stimulated LH release were found between groups even though multiple reproductive experiences resulted in the prolongation of oestrous cyclicity. In Expt 3, day 8 lactating primiparous rats were administered 2, 5, 10 or 25 mg naloxone kg-1 i.v. The three lowest naloxone doses, but not the 25 mg kg-1 dose, significantly increased LH concentrations. The possible effects of prior reproductive experience on opioid control of LH during lactation were then investigated. Naloxone at 0.5 mg kg-1, but not at 2 mg kg-1 or 10 mg kg-1, stimulated a significantly greater rise in LH in multiparous (two litters) than in primiparous females. Overall, these data indicate that while modest differences were found in naloxone-induced LH responses between multiparous and primiparous animals during lactation, reproductive experience did not significantly alter opioid regulation of LH during subsequent oestrous cycles at the naloxone doses examined. Hence, the effects of reproductive experience on opioid regulation of LH are less pronounced than those previously found for opioid regulation of prolactin and behaviour.

Reproductive experience reduces haloperidol-induced prolactin secretion in female rats.

The present study examined possible changes in the dopaminergic regulation of prolactin (PRL) secretion which have been reported to occur in reproductively experienced female rats. In the first experiment, female rats which had raised a litter to weaning and age-matched nulliparous controls were ovariectomized and challenged sequentially 2 days apart with a low (0.2 mg/kg) and high (1.0 mg/kg) dose (intravenously) of haloperidol (HAL), a dopamine antagonist. Blood samples were collected via intra-atrial catheters, and plasma samples were assayed for PRL content by radioimmunoassay. Whereas HAL stimulated increases in plasma PRL concentrations in both nulliparous and primiparous animals, significantly higher levels of plasma PRL were present after both doses of HAL in the nulliparous rats. A second experiment investigated the role of lactation in this change in the ability of HAL to stimulate increases in circulating PRL levels. Separate sets of age-matched primiparous (pups removed on day 1 of lactation) and nulliparous rats were challenged with two doses of HAL 2 weeks after gonadectomy. In contrast to the effect of pregnancy and lactation found in the first experiment, pregnancy and parturition in the absence of lactation failed to alter the female's sensitivity to HAL. The PRL responses in the two groups were identical at the low HAL dose and similar after the high HAL dose. These findings demonstrate that a single prior pregnancy and lactation, but not pregnancy alone, significantly reduce the ability of HAL to elevate circulating plasma PRL concentrations. Decreased circulating PRL levels in reproductively experienced females, therefore, may result in part from increased endogenous dopaminergic activity/tone.

Differential proopiomelanocortin gene expression in the medial basal hypothalamus of rats during pregnancy and lactation.

Hypothalamic proopiomelanocortin (POMC) gene expression was determined using in situ hybridization histochemistry (ISHH) during pregnancy and lactation in rats with and without prior reproduction experience. POMC mRNA levels in the arcuate nucleus were compared between primigravid (first pregnancy) and multigravid (second pregnancy) and primiparous and multiparous lactating rats, and between these groups and age-matched, regularly cycling, nulliparous females in diestrus. Hybridizations were performed using a digoxigenin-labeled riboprobe complementary to 837 bp of the POMC gene. The number of cells expressing POMC mRNA in the arcuate nucleus decreased in primiparous rats on day 12 of lactation when compared with the number of POMC cells in the arcuate nucleus of nulliparous rats in diestrus. In addition, the number of cells expressing POMC mRNA in multigravid animals was significantly less than in the primigravid group on days 7 and 21 of pregnancy, and on day 12 of lactation in primiparous animals. Repeated reproductive experience affected the number of POMC mRNA positive cells; there were fewer cells expressing POMC mRNA in the multigravid females on day 7 of pregnancy and an increase in the number of POMC cells in the multiparous group on day 12 of lactation compared to the primiparous animals. Optical density measurements revealed a significant increase in reaction product in the labeled cells on all days of pregnancy compared with virgin females in diestrus and a significant decrease in reaction product on day 12 of lactation in the multiparous group. The results of the present study indicate that POMC gene expression changes across pregnancy and lactation and that repeated reproductive experience has long-term, possibly permanent, effects on the endogenous opioid system.

Central lactogenic regulation of maternal behavior in rats: steroid dependence, hormone specificity, and behavioral potencies of rat prolactin and rat placental lactogen I.

Adult virgin female rats display maternal behavior when continuously exposed to foster young for 5-6 days. Central infusions of PRL or placental lactogens (PLs) together with systemic treatment of progesterone (P) and estradiol (E2) stimulate maternal behavior in 1-2 days. In the present set of studies, it was asked whether the actions of lactogenic hormones are dependent upon both E2 and P and specific to lactogenic molecules. Moreover, we wanted to know whether central infusions of rat (r) PRL and PLs were equally effective in inducing maternal behavior. In the first study, adult virgin rats were ovariectomized (ovx) and stereotaxically fitted with bilateral cannulas directed at the medial preoptic area (MPOA). Rats were then assigned to one of four groups: P plus E2, blank (B) plus E2, P plus B, and B plus B. P-filled or B capsules were implanted sc on treatment day 1 and removed on day 11, whereas E2 or B capsules were implanted on day 11. All groups were infused with rPRL (40 ng/side) five times from days 11-13 and injected with bromocriptine (CB-154) sc (days 11-17) to suppress endogenous PRL release. Behavioral testing was conducted daily from days 12-17. It was found that exposure to both P and E2 was necessary to induce a fast onset of maternal behavior in PRL-infused females; priming with P or E2 alone in PRL-treated rats failed to stimulate a fast onset of behavior relative to that in nonsteroid-treated controls. In the second experiment to determine the biochemical specificity of PRL's action, adult nulliparous rats were ovx, implanted with bilateral cannulas directed at the MPOA, treated with both P and E2, injected with CB-154, and infused centrally (five times) with 40 ng (per side) of bovine GH, ovine LH, or vehicle. Central infusions of either bovine GH or ovine LH failed to stimulate maternal behavior, suggesting that the stimulatory actions of PRL are related to its lactogenic properties. In the final study, rats were ovx, fitted with bilateral cannulas directed at the MPOA; treated with P, E2, and CB-154; and given a single set of bilateral infusions of rPL-I or rPRL (40 ng/side.infusion) on day 11, three sets of infusions of rPL-I or rPRL (days 11 and 12), or vehicle infusions. Rats given three infusions of rPL-I and rPRL responded faster than controls, although the effect was not as robust as that in animals given five infusions in the initial study. rPL-I and rPRL groups did not differ from one another. Together these studies indicate that 1) both P and E2 are required for lactogenic stimulation of maternal behavior; 2) the stimulatory actions of PRL and rPLs on maternal behavior are related to their lactogenic properties; 3) extended treatment of females with lactogenic hormones is more effective in stimulating the onset of maternal behavior; and 4) the neural potencies of rPRL and rPL-I are similar. These findings provide support for the idea that the induction of maternal behavior is stimulated by the central actions of lactogenic hormones.

Endocrine communication between conceptus and mother: placental lactogen stimulation of maternal behavior.

The possible role of the conceptus in stimulating the onset of maternal behavior through its secretion of placental lactogens and their passage into the brain was investigated in female rats. In the first study, significant mitogenic activity in the Nb2 lymphoma cell bioassay was detected in cerebrospinal fluid (CSF) samples collected by push-pull perfusion from rats on days 12-21 of pregnancy, coincident with the establishment of placental function. In contrast, mitogenic activity was absent from CSF in lactating and gonadectomized, virgin females. In a second study the mitogenic activity in day 12 pregnant samples was neutralized 71% with antibodies to rat placental lactogen-I (rPL-I) and > 90% with a combination of antibodies to rPL-I plus rPL-II. In contrast, activity on day 21 of pregnancy, 1 day prepartum, was reduced by antibodies to rPL-II (> 85%), but not by antibodies to rPL-I, indicating that the predominant lactogen in the CSF prepartum is rPL-II. The behavioral actions of placental secretions were assessed in the third experiment by infusing recombinant rPL-I and purified rPL-II directly into the medial preoptic area of the brain of steroid-primed, nulliparous rats. Latencies to respond maternally to foster young were significantly reduced in rPL-I- and rPL-II-treated rats (2- to 3-day latencies) when compared with latencies in control females (5- to 6-day latencies). Thus, the conceptus through its secretion of rPLs which apparently gain access to the CSF helps to prime the pregnant female's brain to respond maternally at the end of gestation. This endocrine communication between the developing conceptus and pregnant female appears to be an important part of the biological system which helps to establish successful maternal care.

Inhibitory effects of naltrexone on the induction of parental behavior in juvenile rats.

Juvenile rats are rapidly responsive to pups soon after weaning, displaying maternal-like behaviors such as licking, retrieving, grouping, and crouching over pups. As juveniles reach 30 days of age, they become less responsive to pups and show increased latencies to display the same parental behaviors. In light of previous data implicating opiates in the display of ongoing maternal behavior, we administered naltrexone, a long-acting opiate antagonist, beginning 5 and 9 days prior to and continuing throughout the period of behavioral testing, which started at 26 or 30 days of age. Male and female juveniles treated with 10 mg/kg of naltrexone SC for 9 days (days 21 to 29 of age) prior to and during behavioral testing (days 30 to 37) showed longer latencies to retrieve, group, and crouch over pups than did the vehicle-injected controls. These results suggest that opioids may have a stimulatory role in parental behavior during this prepubertal period.

Investigation into the role of cholecystokinin (CCK) in the induction and maintenance of maternal behavior in rats.

The biochemical regulation of maternal behavior has been extensively studied. Cholecystokinin (CCK), a gut peptide that is also present in the brain, recently has been implicated in the onset of maternal behavior in estrogen-primed virgin rats. The objective of the present set of studies was to delineate the role of CCK in the onset (Experiments 1-3) and maintenance (Experiments 4 and 5) of maternal behavior in rats. In the first study intracerebroventricular (i.c.v.) administration of CCK was unable to stimulate the onset of maternal behavior in estrogen-primed virgin rats. Similarly, i.c.v. infusions of CCK into pregnant rats, starting on Day 17 of gestation (Experiment 2), did not advance the onset of maternal behavior. Moreover, when CCK-filled minipumps were implanted intraperitoneally in estrogen-primed virgin rats, the rate of onset of maternal behavior was unaffected (Experiment 3). In contrast, direct infusions of CCK into the MPOA blocked the disruptive effects of beta-endorphin on the maintenance of maternal behavior in postpartum lactating rats (Experiment 4). In addition, proglumide, a CCK receptor antagonist, disrupted maternal behavior in postpartum lactating rats by increasing latencies to retrieve and crouch over the young (Experiment 5). These results support an involvement of CCK in the maintenance, but not the onset, of maternal behavior in rats.

Prolactin-brain interactions in the induction of material behavior in rats.

Most adult female mammals display an immediate onset of maternal care toward their offspring at parturition, whereas the responses of inexperienced, nulliparous females are often less intense or absent. The shift from being a slow or nonresponder in nulliparous females to a rapid responder at parturition in primiparous animals is induced in part by the endocrine changes of pregnancy. This report reviews recent evidence demonstrating a role for prolactin in the stimulation of maternal behavior in the rat. Moreover, new findings are presented that indicate that endogenous rat prolactin acts centrally to stimulate maternal behavior in steroid-primed, nulliparous rats and that the ventromedial hypothalamus in addition to the medial preoptic area are important neural substrates regulating the rapid induction of maternal behavior at parturition.

Prior parity reduces post-coital diurnal and nocturnal prolactin surges in rats.

Mating stimuli received by female rats activate a neuroendocrine mnemonic system which produces daily diurnal and nocturnal prolactin (PRL) surges for the first half of gestation, surges which help maintain corpora lutea function and a viable pregnancy. Since these PRL surges may be regulated in part by endogenous opioids and opioid sensitivity declines as a function of multiple births, we decided to investigate the possibility that prior parity might affect the post-coital diurnal and nocturnal PRL surges, reducing their magnitude and/or occurrence. Age-matched, nulliparous and primiparous rats were mated to males from our colony. On days 5 or 10 of pregnancy females received jugular catheters. Blood samples were collected at regular intervals from 1000 h on day 7 to 1000 h on day 8, and from 1000 h on day 12 to 1000 h on day 13 of gestation in separate sets of multigravid and primigravid rats. Measurement of plasma PRL by radioimmunoassay revealed that prior reproductive experience altered the patterns and levels of plasma PRL. Plasma PRL levels were significantly reduced during both the diurnal and nocturnal surges on days 7-8 in multigravid rats when compared with levels in primigravid rats. No differences in PRL levels were found between primigravid and multigravid groups on days 12 to 13 of gestation. The changes in diurnal and nocturnal PRL surges during early pregnancy indicate that prior parity reduces the subsequent secretion of PRL, possibly by altering the neuroendocrine regulation of this hormone.

Diminished luteinizing hormone release in prenatally stressed male rats after exposure to sexually receptive females.

Prenatally stressed (P-S) males show reductions in male sexual behavior, medial preoptic area volume, and levels of circulating testosterone. We examined the luteinizing hormone (LH) response to the presence of a sexually receptive female, a known index of sexual arousal. Adult male offspring from mothers stressed on days 15-22 of pregnancy (thrice-daily exposures to heat, light, and restraint) were implanted with an intraatrial catheter. Forty-eight h later they were placed into test chambers divided by a wire mesh partition; the catheter was extended outside the chamber. Thirty min later a baseline blood sample (time 0 min) was taken and a sexually receptive female was then placed on the side of the partition opposite the male for 60 min. Blood samples were collected 5, 10, 15, 30, 60 min, and 24 h after introduction of the female. Prenatally stressed males exhibited significantly lower LH levels following exposure to the female at each time point, 5 min to 24 h. Furthermore, at no point were P-S males' LH levels significantly increased above baseline levels. These data suggest that attenuations in female-induced LH release are associated with the marked reductions in male sexual behavior characteristic of the P-S male.

Neural and endocrine sensitivities to opioids decline as a function of multiparity in the rat.

Hormonal changes during pregnancy regulate the onset of maternal behavior at parturition. In addition, the concentrations of beta-endorphin and mu opioid receptors are higher during pregnancy and lower during lactation. Previous studies have shown that sensitivity of female rats to the disruptive behavioral effects of morphine changes as a function of the number of pregnancies and/or lactations the females undergo. The objectives of the present study were to determine whether central infusions of the endogenous opioid, beta-endorphin, would disrupt maternal behavior. Next, we investigated the possibility that the neural sensitivity to beta-endorphin changes with repeated pregnancies. And finally, we examined whether opioid-mediated endocrine responses also change as a function of multiparity. In the first study, bilateral infusions of low doses (0.06-0.72 nmol) of beta-endorphin into the medial preoptic area (MPOA) of lactating, primiparous rats disrupted maternal behavior. When comparable doses of beta-endorphin were infused into the MPOA of age-matched, multiparous rats, the behavioral effects of beta-endorphin were significantly attenuated. In response to suckling stimulation, an opioid-mediated endocrine response, primiparous mothers secreted more prolactin than did multiparous rats. Moreover, multiparous, but not primiparous, mothers were insensitive to the ability of naloxone, an opiate antagonist, to block suckling-induced increases in prolactin. These findings indicate that reductions in neural sensitivity to opioids develop as females undergo repeated pregnancies and lactations, changes which affect both behavioral and endocrine functions.

Intracerebroventricular cholecystokinin infusions block beta-endorphin-induced disruption of maternal behavior.

Recent work has shown that infusions of beta-endorphin, an endogenous opioid, into the ventricular system of lactating rats blocks normal maternal behavior. Other behavioral and biochemical studies have demonstrated that sulfated cholecystokinin-octapeptide (CCK-8) can have effects opposite those of opioids. The present study evaluated whether intracerebroventricular (ICV) administration of CCK-8 is able to antagonize the inhibitory effect of beta-endorphin on maternal behavior. The results of this study demonstrated that CCK-8 (14.5 nmol) prevented the beta-endorphin (1.45 nmol)-induced increase in latencies to retrieve the first pup, retrieve all pups, and to group and crouch over rat pups. In addition, reductions in the percentage of rats retrieving all pups and displaying full maternal behavior were prevented by CCK-8. These data suggest that CCK-8 can act as an opioid antagonist in neural systems that control maternal behavior.

Opioid receptor subtype involvement in maternal behavior in lactating rats.

Central or systemic administration of morphine disrupts maternal behavior in steroid-primed, pup-induced virgin and lactating rats. Morphine, the prototypical mu agonist, also interacts with different opioid receptor subtypes. The present study examined the effectiveness of five receptor-selective agonists, in addition to morphine, to disrupt maternal behavior in primiparous lactating rats following intracerebroventricular (i.c.v.) infusions in order to characterize opioid receptor subtype involvement in maternal behavior in the female rat. Virgin, Sprague-Dawley rats were mated and implanted with lateral ventricle cannulae on days 13-15 of gestation. On postpartum day 5, mothers were tested for maternal behavior 30 min after i.c.v. vehicle infusion (5 microliters). On day 6, rats received one of the following opioid receptor agonists 30 min before testing: beta-endorphin (mu/epsilon receptor subtype; 0.29, 0.72, 1.45, 2.9 nmol), DAGO (mu; 0.29, 0.72, 1.45, 2.9 nmol), morphine (mu; 0.29, 0.72, 1.45, 2.9, 14.5 nmol), DPDPE (delta; 2.9, 29 nmol), U50488H (kappa l; 2.9, 29, 145 nmol) and SKF10047 (sigma; 2.9, 29, 145 nmol). Only activation of mu opioid receptors dose-dependently disrupted maternal behavior in primiparous lactating rats. DPDPE, U50488 and SKF10047 had no discernible effect on maternal behavior. DAGO, a highly selective mu agonist, was even more potent than beta-endorphin and morphine in disrupting maternal behavior suggesting that maternal behavior is regulated by opioids interacting with the mu opioid receptor.

Central prolactin infusions stimulate maternal behavior in steroid-treated, nulliparous female rats.

A series of experiments were conducted to determine whether and under what conditions central prolactin (PRL) administration would stimulate the onset of maternal behavior in female rats and to identify possible neural sites of PRL action. In each experiment ovariectomized, nulliparous rats whose endogenous PRL levels were suppressed with bromocriptine were tested for maternal behavior toward foster young. In experiments 1, 2, and 4, females were also exposed to pregnancy-like levels of progesterone (days 1-11) followed by estradiol (days 11-17). In experiment 1 infusions (days 11-13) of four doses of ovine PRL (400 ng, 2 micrograms, 10 micrograms, or 50 micrograms, but not 80 ng) into the lateral ventricle resulted in a rapid onset of maternal behavior (behavioral testing, days 12-17). The stimulatory action of these doses of PRL appears to be central, since subcutaneous injections of 50 micrograms of ovine PRL failed to affect maternal responsiveness (experiment 2). Experiment 3 indicated that the stimulatory effect of intracerebroventricularly administered PRL is steroid dependent. Infusions of either 10 micrograms of ovine PRL or 10 micrograms of rat PRL failed to induce maternal behavior in nonsteroid-treated animals. In the final experiment (no. 4) bilateral infusions of 40 ng of ovine PRL into the medial preoptic area of steroid-treated rats resulted in a pronounced stimulation of maternal behavior. These findings demonstrate a central site of PRL action in the stimulation of maternal responsiveness and point to the medial preoptic area as a key neural site for PRL regulation of maternal behavior.

Prenatal stress has long-term effects on brain opiate receptors.

Prenatal stress has been associated with a number of behavioral consequences including altered sensitivity to exogenous opiates. In the present study, mu opiate receptors were compared in the 42-day-old offspring from females stressed on days 15-22 of gestation and from females who were unstressed controls. Membrane homogenates from the prenatal stress group showed less binding of the mu opiate receptor ligand, [3H]DAGO in striatum but not in several other brain regions. Saturation studies suggest this difference is due to fewer striatal mu opiate receptors in offspring of prenatally stressed females. Using in vitro receptor autoradiography, the decreased binding in striatum was found mostly in the rostral striatum, extending into the nucleus accumbens with conservation of the normal anatomic distribution of receptor rich patches.

Antagonism of morphine analgesia by nonopioid cold-water swim analgesia: direct evidence for collateral inhibition.

The demonstrated existence of opioid and nonopioid forms of pain control has raised questions as to how they interact. Previous indirect evidence suggests that activation of one system inhibited the activation of the other. The present study assessed this directly using morphine as an opiate form of analgesia and continuous cold-water swims (CCWS, 4 degrees C, 2 min) as the nonopioid form. A significant reduction in morphine (8 mg/kg, SC) analgesia on the tail-flick test was observed if rats were acutely exposed to CCWS immediately prior to morphine administration. The inability of naloxone (10 mg/kg, SC) to reduce CCWS analgesia verified its nonopioid nature. The antagonism of morphine (3 mg/kg, SC) analgesia was greater following preexposure to 2 min of CCWS than 1 min of CCWS. CCWS was also more effective in antagonizing analgesia induced by the 3 mg/kg than the 8 mg/kg dose of morphine. The antagonism of morphine analgesia by CCWS was dependent upon the temporal patterning of stimulus presentation: exposure to CCWS 20 or 60 min prior to morphine failed to alter subsequent morphine analgesia. A significant reduction in analgesia induced by intraperitoneal administration of morphine (10 mg/kg) was also observed when CCWS was presented immediately prior to injection, suggesting that pharmacokinetic factors such as altered drug absorbance by CCWS-induced vasoconstriction do not appear to explain these effects. These data provide direct support for the existence of collateral inhibitory mechanisms activated by CCWS and morphine, and suggests that these opioid and nonopioid forms of analgesia do not function synergistically, but instead involve some form of hierarchical order.