Estrogen receptor-alpha knockouts and maternal memory in nulliparous rats.

The current study investigated the role of estrogen receptor alpha (Esr1) in maternal memory in rats, comparing the induction and retention responses of Esr1 knockout (KO) and wild type (WT) nulliparous rats towards foster pups. Thirty days after completion of induction testing, subjects were tested for the retention of maternal care in their home cage and then for maternal behaviors in a novel cage. Both WT and Esr1 KO females displayed similar latencies to respond to foster young during the initial induction testing. Likewise, reinduction latencies to display full maternal responsiveness were similar in the Esr1 KO and WT groups during maternal memory testing in the home cage. However, in the novel cage testing WT subjects displayed modest modifications in maternal care. WT females had shorter latencies to first retrieve and mouth a test pup. These findings suggest that while Esr1 does not appear to affect the establishment of maternal care or the display of maternal memory, it may modulate aspects of pup-directed behaviors associated with the reinduction of maternal care in female rats.

Reversal of morphine disruption of maternal behavior by concurrent treatment with the opiate antagonist naloxone.

Rats whose pregnancies were surgically terminated on day 17 of gestation were injected with morphine, morphine plus naloxone hydrochloride, or saline, and then tested for maternal responsiveness toward foster young. Morphine treatment alone significantly disrupted the rate of onset and quality of maternal responsiveness. Concurrent administration of naloxone to morphine-injected rats reinstated the rapid onset of behavioral responsiveness toward foster young, such that the responsiveness of the rats treated with both morphine and naloxone was indistinguishable from that shown by saline-injected controls. The disruptive effects of morphine did not appear to result from a general reduction in activity levels as measured in an open-field apparatus. These findings suggest that the normal onset and maintenance of maternal behavior in the rat may be regulated by endogenous opiates.

Prolactin stimulation of maternal behavior in female rats.

Inexperienced, hypophysectomized female rats treated with steroids were used in experiments to investigate the roles of the pituitary gland and prolactin in the expression of maternal behavior. Administration of ovine prolactin or treatment with ectopic pituitary grafts, which release prolactin into the circulation, stimulated maternal care in these females toward rat young. Steroid treatment alone, while stimulating maternal behavior in rats with intact pituitary glands, did not facilitate maternal responsiveness in hypophysectomized females. These findings indicate a stimulatory behavioral role for pituitary prolactin in the establishment of maternal care and suggest that exposure to prolactin during pregnancy helps to stimulate the immediate onset of maternal behavior at parturition.

Reproductive experience and expression of dopamine D(2) receptor mRNA: a possible mechanism for reduced prolactin secretion in primiparous rats.

Reproductive experience (i.e. pregnancy and lactation) leads to reduced levels of circulating prolactin in both women and rats. Stimulation of prolactin secretion by dopamine antagonists is also blunted following reproductive experience in both species. Whereas a parity-induced reduction in haloperidol-stimulated prolactin secretion is evident in ovariectomised rats, it is unknown whether a similar attenuation of prolactin secretion is present in reproductively experienced, cycling pro-oestrous rats. The present study examined this possibility. Moreover, to determine possible mechanisms involved in parity-mediated changes in prolactin secretion, both dopamine utilisation within the arcuate nucleus/median eminence and expression of dopamine D(2) receptor mRNA (short and long forms) in the anterior pituitary were measured across the afternoon of pro-oestrous in reproductively experience and inexperienced females. Prolactin secretion was lower on the afternoon of pro-oestrous in primiparous females compared to age-matched, nulliparous controls. In addition, haloperidol-stimulated prolactin secretion was reduced in ovariectomised, reproductively experienced females. Although no differences in dopamine utilisation were observed as a function of reproductive experience, parity did affect the expression of both forms of D(2) receptor mRNA in the anterior pituitary. Compared with nulliparous controls, primiparous females had increased D(2 long) mRNA expression at 12.00 h on pro-oestrous as well as increased D(2 short) mRNA expression at 14.00 h. Because the ratio of D(2 long)/D(2 short) can significantly effect lactotroph proliferation and prolactin secretion, a shift in relative expression of the two D(2) receptor isoforms within the anterior pituitary of parous females may help account for the reduction in prolactin secretion that occurs following reproductive experience.

A quantitative analysis of the roles of dosage, sequence, and duration of estradiol and progesterone exposure in the regulation of maternal behavior in the rat.

The regulation of the onset of maternal behavior in the rat is under hormonal control. This study reports a new endocrine model for the study of the hormonal regulation of maternal responsiveness. The model employs the administration of physiological amounts of the steroids estradiol (E2) and progesterone (P) via Silastic implants to inexperienced nulliparous rats and measurement of the effects of these implants on maternal behavior. In the first two experiments, the levels of E2 and P in the sera of pregnant and hormone-treated rats were measured by RIA. Using known physiological treatments of E2 given in combination with P, the effects of E2 and P on maternal behavior were measured. Treatment with a combination of E2 at all dosages plus P for 2 weeks before P removal and behavioral testing stimulated a fast onset of maternal behavior in ovariectomized nulliparous rats. Exposure for 2 weeks to small E2 implants (1 or 2 mm; approximately 20-30 pg/ml serum) did not affect maternal responsiveness, whereas large E2 implants (10 mm; approximately 110 pg/ml serum) stimulated maternal behavior. P treatment alone had no behavioral effect. Simultaneous removal of E2 plus P before exposure to foster young also resulted in a stimulation of behavioral responsiveness, indicating that the presence of elevated titers of circulating E2 is not a requirement for stimulation to occur. In addition to facilitating a rapid onset of behavior, the quality of the response in steroid-primed rats was similar to that measured in lactating rats in a T-maze test. In another experiment, when female rats were treated with P before E2 administration, maternal behavior was rapidly induced. Thus, P itself can sensitize the female to the behavioral effects of E2. Finally, the duration of steroid-exposure before testing was found to influence maternal behavior. Increased durations of E2 plus P exposure before testing were accompanied by decreased latencies to respond maternally to foster young. These data indicate that during pregnancy, E2 and P prime the female to respond to her young at birth. The intensity of the steroidal priming increases as pregnancy progresses, and this primed potential is subsequently unmasked by the decline in P and the maintenance of E2 secretion around parturition. These findings demonstrate that behavioral processes can be modified in the adult animal as a result of long term changes in endocrine state, i.e. pregnancy.

Ovarian control of prolactin secretion during late pregnancy in the rat.

The involvement of the ovaries during late pregnancy in the rat upon serum prolactin was investigated. Ovariectomy on day 17 or day 21 of pregnancy prevented the dramatic rise of prolactin found in sham-ovariectomized animals between days 21 and 23 of pregnancy. While animals ovariectomized on day 17 of pregnancy failed to carry viable fetuses beyond day 19 of pregnancy, rats ovariectomized on day 21 of pregnancy had viable pups in utero on day 23 of pregnancy. These data suggest that the rise in serum prolactin during late pregnancy is stimulated by ovarian factors.

Plasma prolactin concentrations in parental male and female rats: effects of exposure to rat young.

The effects of pup presentation on the PRL responses in parental male rats were measured and compared with those in parental virgin and lactating female rats. Blood samples were collected from rats through indwelling intraatrial cannulas after a suckling challenge, i.e. presentation of rat young. Lactating rats showed full parental behavior and characteristic large surges in plasma PRL levels within the first 5-10 min on each day that rat young were presented (days 4, 8, and 12 of lactation). When pups were not presented, PRL rises did not occur. In contrast to the pattern of PRL responses shown by lactating mothers, parental ovariectomized nulliparous female and parental intact male rats failed to show specific increases in PRL in response to pup presentation. Plasma PRL levels in these groups, as in nonparental female and male rats, occasionally rose in response to blood collection rather than to pup presentation alone. Treatment of nulliparous female as well as male rats with estradiol and progesterone Silastic implants for 21 days before the initiation of behavioral testing significantly reduced the latencies of both nulliparous females and males to respond to foster young from about 5 to 2 days. The PRL responses of these steroid-primed groups were quite different. The steroid-primed females exhibited a pattern of PRL responses to pups identical to that found in lactating rats. The steroid-primed parental males, in contrast, failed to show specific increases in plasma PRL levels in response to young. These data demonstrate a sex difference in the hormonal, but not behavioral, responses of male and female rats to young and are suggestive of possible sex differences in the hypothalamic and/or peripheral regulation of pup-induced PRL secretion.

Prolactin (PRL) regulation of maternal behavior in rats: bromocriptine treatment delays and PRL promotes the rapid onset of behavior.

Recent findings indicate that PRL helps stimulate the onset of maternal behavior in inexperienced hypophysectomized steroid-treated female rats. In a series of five experiments we have further examined the involvement of PRL in maternal behavior using nonhypophysectomized ovariectomized rats treated concurrently (type I) or sequentially (type II) with progesterone (P) and estradiol (E2) and administered either bromocriptine (to suppress endogenous PRL secretion) or bromocriptine plus ovine PRL. In Exp 1 plasma PRL concentrations were measured in ovariectomized rats treated for 2 weeks with a combination of E2 and P Silastic implants. Type I steroid-treated (2mm E2, days 1-24; three 30 mm P, days 3-13) rats exhibited elevated plasma PRL levels throughout the sampling period compared with nonsteroid-treated controls. In contrast, PRL concentrations in type II steroid-treated (P, days 3-13; E2, days 13-24) females were low (similar to controls) from days 3-13 when the type II steroid-treated females were exposed to P only. Like type I treated rats, PRL levels in type II steroid-treated rats were elevated from day 13 onward after E2 capsule insertion. In Exp 2, treatment of both type I and type II steroid-treated rats with bromocriptine (2 mg/kg, sc) twice daily beginning on treatment day 13 suppressed basal PRL concentrations and prevented the estrogen-induced diurnal PRL surge. Whereas PRL was effectively suppressed by bromocriptine in both steroid-treated groups, the absolute levels of PRL were lower in rats treated with the type II steroid regimen. Behavioral analyses in Exp 3, 4, and 5 revealed that bromocriptine administration, while failing to interfere with the onset of maternal behavior in rats treated with the type I concurrent steroid regimen, disrupted the onset of maternal care in rats treated with the type II sequential steroid regimen. When a separate set of type II steroid-treated rats was given both bromocriptine (2 mg/kg) plus ovine PRL (0.5 mg, sc) twice daily, maternal behavior rapidly appeared. Thus, suppression of endogenous PRL secretion delays the onset of maternal behavior in nonhypophysectomized steroid-primed rats, an effect prevented by concurrent administration of ovine PRL. In addition to providing further experimental support for PRL's role in maternal behavior, the development of this endocrine regimen provides researchers with a potentially fruitful model to examine neural sites and mechanisms of PRL regulation of maternal behavior in mammals.

Null mutation of the prolactin receptor gene produces a defect in maternal behavior.

We have studied pup-directed maternal behavior in mice carrying a germ line null mutation of the PRL receptor (PRLR) gene. Homozygous mutant and heterozygous mutant nulliparous females show a deficiency in pup-induced maternal behavior. Moreover, primiparous heterozygous females exhibit a profound deficit in maternal care when challenged with foster pups. Morris maze studies revealed normal configural learning in the heterozygous and homozygous animals. Eating, locomotor activity, sexual behavior, and exploration (all processes regulated by the hypothalamus) are normal in PRLR mutant mice. Olfactory function was tested in an aversive conditioning paradigm, results indicating that heterozygous and homozygous PRLR mutant mice are not anosmic. These studies clearly establish the PRLR as a regulator of maternal behavior.

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.

Hypothalamic LHRH in aging rats: effects of ovariectomy and steroid replacement.

Hypothalamic LHRH was measured by RIA in young and middle-aged (MA) female rats in several endocrine conditions. Temporal alterations in LHRH content associated with the steroid induced gonadotropin surge were compared in medial basal hypothalamic and anterior hypothalamic-preoptic area fragments of ovariectomized young and MA subjects. LHRH content was also compared in ovariectomized, untreated subjects from the two age groups. Finally, LHRH content in MA constant estrous females was compared with content in young females on the morning of proestrus. In all conditions, LHRH levels in both brain regions of MA females were similar to, or significantly elevated above levels measured in young females, yet both the steroid induced surge and the castration induced hypersecretion of gonadotropins were markedly attenuated in aging females. Because studies have verified the responsiveness of the pituitary of MA rats to LHRH, the data suggest that adequate amounts of hypothalamic LHRH do not reach the pituitary. Rather, high levels of hypothalamic LHRH measured in MA subjects may represent accumulation of the peptide in LHRH neurons due to an age-related impairment in release.

Steroidal interactions in the regulation of maternal behaviour in virgin female rats: effects of testosterone, dihydrotestosterone, oestradiol, progesterone and the aromatase inhibitor, 1,4,6-androstatriene-3,17-dione.

The effects of exposure to concentrations of androgens, oestradiol (OE2) and progesterone similar to those found during pregnancy on the induction of maternal behaviour were investigated in female rats. In the first experiment the effects of testosterone and dihydrotestosterone (DHT) administered in combination with progesterone (using silicone elastomer capsules) on the induction of behavioural responsiveness towards young (crouching, retrieval and grouping of pups) were measured in ovariectomized virgin rats. Hormonally treated animals were exposed to testosterone or DHT from day 1 of treatment to the end of behavioural testing, while progesterone was administered for 10 days (days 3-13). Testing for maternal responsiveness began on day 14 and lasted until day 23. Significant reductions in latencies to show individual aspects of and complete maternal behaviour were found only in animals treated with a combination of testosterone and progesterone (range of mean latencies for showing one aspect of, to complete, maternal behaviour = 1.0-1.4 days). The mean latencies of the other hormonally treated animals ranged from 5 to 6 days and were similar to those of non-hormonally treated control rats. The second experiment examined the possibility that stimulation of maternal behaviour in animals given testosterone and progesterone resulted from the aromatization of testosterone to OE2. Ovariectomized virgin rats were implanted with capsules containing testosterone and other with the aromatase inhibitor, 1,4,6-androstatriene-3,17-dione (ATD) on day 1, and with progesterone capsules on day 3. Progesterone capsules were removed on day 13 and behavioural testing commenced on day 14. Treatment with testosterone and progesterone failed to stimulate maternal behaviour in rats treated with ATD. In a third study ovariectomized virgin rats were implanted with OE2 on day 1 and progesterone on day 3. The progesterone implants were removed on day 13 and testing began on day 14. Significant reductions in latencies to show all aspects of maternal behaviour were found in these rats. In a final study progesterone capsules remained in OE2- and progesterone-treated rats from day 3 until the completion of behavioural testing. The presence of progesterone implants throughout the test period (days 14-23) blocked the rapid onset of maternal responsiveness induced by removal of progesterone on day 13 shown in rats treated with OE2 plus progesterone in experiment 3. These data suggest that during gestation testosterone, through its conversion to OE2, synergizes with progesterone to help stimulate the development of the capacity of the female animal to respond maternally to young, a capacity unmasked by withdrawal of progesterone before parturition.

Sex differences in prolactin secretion in parental male and female rats.

The present series of experiments report the behavioral and endocrine responses of male and female rats to rat young. In the first experiment, the behavioral responses of intact male and ovariectomized female rats to foster young were measured. While both of these groups exhibited most components of parental behavior after four to six days of exposure to rat young, males failed to build good nests and to crouch over the young. In addition significantly more males killed young than did ovariectomized females. In the second experiment, treatment of male and virgin female rats with a 21-day hormone regimen of estradiol and progesterone stimulated an equally rapid onset of parental behavior in both sexes. Latencies to exhibit parental responses were approx. two days for both males and females. Pup killing was virtually eliminated in males by hormone treatment. Although the behavioral responses of hormone-primed males and females were similar, their prolactin responses to the presentation of rat young differed. Pup presentation stimulated an increase in plasma prolactin content in hormone-primed parental virgin females similar, but not identical, to that found in lactating control rats, while plasma prolactin levels in hormone-primed parental males did not rise in response to the presentation of young. These data indicate that a sex difference in neuroendocrine, but not behavioral, responsiveness to young exists between parental males and females.

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.

Regulation of prolactin secretion in parental rats: roles of steroid priming and pituitary responsiveness.

Whereas prolactin (PRL) release increases in female rats as a result of stimuli from rat young, parental male rats fail to show a rise in plasma PRL levels in response to pup stimuli. In the present study, we have characterized two aspects of the endocrine control of PRL release in male and female rats in an attempt to account for this sex difference in pup-induced PRL secretion. First, we compared the patterns of PRL secretion throughout a prolonged steroid priming treatment that effectively stimulates the onset of parental responsiveness in both sexes. Second, we measured pituitary responsiveness to a PRL releasing agent (thyrotropin releasing hormone: TRH) in male and female rats that had previously experienced the steroid treatment. We found that in the presence of estradiol alone or soon after the onset of progesterone treatment in the presence of estrogen, female but not male rats displayed afternoon elevations in PRL. We also found that, following steroid treatment, TRH caused significant and comparable elevations in plasma PRL in both sexes. These experiments demonstrate differences in patterns of PRL secretion in male and female rats during the early phase of a steroid priming treatment that induces parental responsiveness, yet comparable pituitary sensitivity to TRH-induced PRL release following this 3 week steroid treatment. These data suggest that sex differences in pup-induced PRL secretion in rats do not result from fundamental differences in endocrine priming or from sex differences in pituitary responsiveness.

Nipple development and pup-induced prolactin release in male rats treated prenatally with the antiandrogen flutamide.

While maternal female rats display increases in circulating prolactin (PRL) concentrations in response to pup exposure, parental male rats fail to show such an increase. One possible explanation for the lack of an acute PRL response in parental male rats is that males do not have nipples and therefore do not receive stimuli from the pups comparable to those experienced by parental female rats. To examine the contribution of nipple presence and possible stimulation, i.e. suckling, in this sexually differentiated endocrine response, male rats were exposed from days 12-15 of gestation to the antiandrogen flutamide. As adults, flutamide-exposed males had nipples. These males and a group of control males were castrated in adulthood and treated with a 21-day hormone regimen (estradiol and progesterone) that effectively stimulates parental behavior in adult rats. Following hormone treatment, mammary tissue from one set of flutamide-treated males was examined histologically to assess nipple and mammary gland development and responsiveness of these tissues to hormonal stimulation. Additional sets of flutamide-treated and control animals were tested for parental behavior. These latter animals were implanted with indwelling atrial cannulae on day 4 of parental behavior and subsequently bled on day 6 in the absence of young and on day 8 after presentation of young. PRL concentrations did not change in either the flutamide-treated or control parental males bled in the presence or absence of young.(ABSTRACT TRUNCATED AT 250 WORDS)

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.

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.

Immunoreactive Prolactin in the Cerebrospinal Fluid of Estrogen-Treated and Lactating Rats as Determined by Push-pull Perfusion of the Lateral Ventricles.

Abstract Immunoreactive (ir) prolactin levels in the cerebrospinal fluid were measured in push-pull perfusion samples collected from the lateral cerebral ventricles in freely-moving ovariectomized rats prior to and after estrogen treatment, and in rats during pregnancy and lactation. Ir-prolactin levels in cerebrospinal fluid were elevated throughout the perfusion period in ovariectomized animals given estrogen and in Day 8 lactating females following the onset of nursing by their young. Cerebrospinal fluid perfusates collected from ovariectomized animals prior to estradiol treatment and from females on Days 18 and 22 of gestation contained levels of ir-prolactin that were below the limit of detectability of the prolactin radioimmunoassay. Data from previous studies in our laboratory suggest that pituitary prolactin is involved in the mediation of maternal responsiveness. Where and how prolactin exerts its actions in this regard remains to be determined. Results of the present study indicate that detectable levels of ir-prolactin are present in the cerebrospinal fluid of nulliparous females after exposure to an estradiol regimen known to shorten their latency to display maternal responsiveness and in lactating females actively exhibiting maternal behavior. Once in the cerebrospinal fluid, prolactin should have access to neural target sites behind the blood-brain barrier. The presence of ir-prolactin in the cerebrospinal fluid of females in the present study is consistent with a role for this protein hormone, acting at the level of the central nervous system, in the facilitation of maternal responsiveness as well as in the regulation of other neural and neuroendocrine processes.