Prolactin receptors in the brain during pregnancy and lactation: implications for behavior

Numerous studies have documented prolactin regulation of a variety of brain functions, including maternal behavior, regulation of oxytocin neurons, regulation of feeding and appetite, suppression of ACTH secretion in response to stress, and suppression of fertility. We have observed marked changes in expression of prolactin receptors in specific hypothalamic nuclei during pregnancy and lactation. This has important implications for neuronal functions regulated by prolactin. In light of the high circulating levels of prolactin during pregnancy and lactation and the increased expression of prolactin receptors in the hypothalamus, many of these functions may be enhanced or exaggerated in the maternal brain. The adaptations of the maternal brain allow the female to exhibit the appropriate behavior to feed and nurture her offspring, to adjust to the nutritional and metabolic demands of milk production, and to maintain appropriate hormone secretion to allow milk synthesis, secretion, and ejection. This review aims to summarize the evidence that prolactin plays a key role in regulating hypothalamic function during lactation and to discuss the hypothesis that the overall role of prolactin is to organize and coordinate this wide range of behavioral and neuroendocrine adaptations during pregnancy and lactation.     

Termination of pregnancy after accelerated lactation in the rat. III. Effects of hormonal treatments.

The relationship was investigated of the hormones associated with pregnancy and lactation and the termination of pregnancy that occurs when rats mated at the post-partum oestrus are allowed to suckle a large litter after implantation. The primary cause for pregnancy termination was found to be an insufficient level of progesterone with the possible need for a primary or synergistic dose of oestrogen. The progesterone deficiency was not due to the high levels of prolactin present in nursing rats, since pregnancy termination could not be produced by prolactin administration in the absence of accelerated lactation, and could not be prevented by blocking prolactin secretion after accelerated lactation. Administration of LH to rats after accelerated lactation did prevent termination of pregnancy.     

Role of milk secretion in transport of prolactin from blood into milk

In a first experiment prolactin was shown to be uniformly distributed in the milk of dairy cows. Accumulation of prolactin in milk increased with milking interval although milk prolactin content was proportionally greater after 2 h than accumulation of milk. Concentration of milk prolactin and total transfer of prolactin into milk was greater in mammary glands of cows milked just prior to initiation of intravenous infusion of exogenous prolactin. Similarly, inhibition of milk secretion as induced by intramammary infusion of colchicine, markedly reduced milk prolactin concentration and total transfer of prolactin into milk 5h following start of intravenous infusion of exogenous prolactin. Data indicate that transfer of prolactin from blood into milk during established lactation occurs primarily in conjunction with milk secretion.     

Aspects of the biochemistry, physiology and endocrinology of lactation

Recent research on the endocrine control of the initiation of lactation, on the hypothalamic control of prolactin secretion, and on the utilization of nutrients by the mammary gland for the synthesis of milk is reviewed. Particular emphasis is placed on the mechanism of action of prolactin, and the role played by prolactin receptors.     

Lactation reduces prolactin levels in reproductively experienced female rats

Long-term alterations in prolactin (PRL) secretion following reproductive experience have been demonstrated in both women and female rats. In the rat, these changes include decreased PRL secretion in response to a dopamine antagonist challenge following ovariectomy, decreased post-coital diurnal and nocturnal prolactin surges in multigravid versus primigravid females, as well as decreased suckling-induced prolactin release in multiparous versus primiparous females. To date, there have been no studies examining PRL secretion following reproductive experience in cycling female rats. Studies in women, however, have demonstrated a reduction in basal PRL secretion during the menstrual cycle. The purpose of the present work was to determine whether similar changes occur in the rat during the estrous cycle and to what extent lactation is involved in these effects. In addition to examining PRL, potential parity-induced changes in estradiol secretion were also studied. The findings revealed a significant decrease in PRL levels during the afternoon of proestrus, which was only observed in primiparous females that had lactated. Significant differences in estradiol secretion were not detected following reproductive experience. Thus, a reduction in the PRL surge on the afternoon of proestrus is a consequence of reproductive experience that requires both pregnancy and lactation.     

The endocrine regulation of milk lipid synthesis and secretion in tammar wallaby (Macropus eugenii)

Lipids in tammar milk are predominantly triacylglycerols, and the fatty acid composition varies during the lactation cycle. Little is known about the regulation of their synthesis. This study investigates the endocrine regulation of lipid synthesis in mammary explants from pregnant tammars. Treatment of mammary explants with insulin resulted in a high level of lipid synthesis, but the lipids accumulated in the cytosol. Culture with prolactin resulted in a small increase in lipid synthesis, but electron microscopy showed lipid globules were synthesized in the mammary epithelial cells and secreted into the lumen. Culture with both insulin and prolactin demonstrated elevated levels of synthesis and secretion of lipid. Analysis of the type of fatty acids synthesized in these mammary explants showed that the initiation of synthesis of C(16:0), which also occurs in the first week of lactation, could be reproduced in the pregnant explants cultured with prolactin alone. However, treatment of mammary explants with hydrocortisone did not show a significant effect on lipid synthesis, secretion or the fatty acid synthesized. These results provide new information identifying the role of insulin and prolactin in regulating milk lipid synthesis and secretion in the tammar.     

The endocrine regulation of milk lipid synthesis and secretion in tammar wallaby (Macropus eugenii)

Lipids in tammar milk are predominantly triacylglycerols, and the fatty acid composition varies during the lactation cycle. Little is known about the regulation of their synthesis. This study investigates the endocrine regulation of lipid synthesis in mammary explants from pregnant tammars. Treatment of mammary explants with insulin resulted in a high level of lipid synthesis, but the lipids accumulated in the cytosol. Culture with prolactin resulted in a small increase in lipid synthesis, but electron microscopy showed lipid globules were synthesized in the mammary epithelial cells and secreted into the lumen. Culture with both insulin and prolactin demonstrated elevated levels of synthesis and secretion of lipid. Analysis of the type of fatty acids synthesized in these mammary explants showed that the initiation of synthesis of C_16:0, which also occurs in the first week of lactation, could be reproduced in the pregnant explants cultured with prolactin alone. However, treatment of mammary explants with hydrocortisone did not show a significant effect on lipid synthesis, secretion or the fatty acid synthesized. These results provide new information identifying the role of insulin and prolactin in regulating milk lipid synthesis and secretion in the tammar.     

Glycosylated prolactin in the murine pituitary: detection by a novel assay and alteration of concentrations by physiological and pharmacological stimuli

In both rat and mouse pituitary extracts, we detected concanavalin A-binding prolactin immunoreactivity by a lectin-binding radioimmunoassay developed recently. The activity increased in response to estradiol benzoate treatment and lactation, stimuli that augment prolactin secretion, and decreased in response to acute nursing and perphenzine administration, stimuli that cause massive release of prolactin. Western blot analysis revealed a prolactin-immunoreactive band 2,000-3,000 greater in Mr than the main prolactin band that bound to 125I-labeled concanavalin A. These results suggest the existence in the murine adenohypophysis of a glycosylated form of prolactin, which seems to be released under certain physiological states.     

Cytological and immunocytochemical studies of the anterior pituitary gland of the beagle bitch during various reproduction phases]

The changes in anterior pituitary (AP) of pregnant and lactating dogs as compared with pituitary of animals in metestrus-anestrus phase are described with special reference to the relative proportion, topography and morphology of prolactin cells, somatotrophs, corticotrophs, thyrotrophs and gonadotrophs. For demonstration of these cells, suitable histological, histochemical and immunoenzyme cytochemical methods are used. The prolactin cells show progressive hyperplasia during pregnancy, so that at the end of this phase and during lactation, they comprise the most predominant glandular cells in AP. At the same time, they reveal massive hypertrophy with marked morphological features of high secretory activity, After transient or continous interruption of the suckling stimulus they show signs of functional inhibition on involution. The corticotrophs appear at 20. and 30. days of pregnancy to be relatively increased in number. While in the last third of pregnancy and during lactation, they only seem to be more active than those in pituitary of metestrus-anestrus dogs. The somatotrophs appear to be progressively reduced in relative number during pregnancy and lactation. However, they show some morphological signs of active secretion. The thyrotrophs did not show any morphological alterations during the different reproductive phases. The gonadotrophs reveal during pregnancy, especially at 30. day morphological signs of stimulation. On the contrary they appear atrophied during lactation. This may be a result of suckling stimulus and morphological expression of the inverse relationship in the secretion pattern of gonadotrophins and prolactin in dogs during suckling. The estrogen and progesterone levels in plasma as well as the changes in their relative concentrations may largely account for the structural changes on AP of pregnant dogs. However, neuroendocrine reflexes (e.g. suckling stimulus) seem to be of a great importance for the maintainance of stimulation of prolactin cells during lactation.     

Treatment of lactating sows with the dopamine agonist Cabergoline: effects on LH and prolactin secretion and responses to challenges with naloxone and morphine

This study was conducted to examine the effects of chronic administration of a long-acting dopamine agonist, Cabergoline, on LH and prolactin secretion during lactation in the sow. The effect of the administration of the opioid antagonist naloxone and the agonist morphine in Cabergoline treated animals was also evaluated. In Part I of the experiment, 16 sows were treated as either CONT sows (n=4; control, no treatment); CAB sows (n=4; treated with Cabergoline from days 10 to 26 of lactation); CAB+NAL sows (n=4; received Cabergoline treatment and naloxone challenges); CAB+MORP sows (n=4; treated with Cabergoline and morphine challenges). Plasma LH and prolactin concentrations were measured in blood samples taken from all sows during 6-h periods at days 12, 19 and 26 of lactation. To extend the results at the most critical response period at day 26, another 11 sows were allocated in Part II to either Control (n=3), Cabergoline (n=4) or Cabergoline and morphine (n=4) treatments as for Part I, but the effect of treatments were only confirmed in a single period of sampling at day 26 of lactation. Cabergoline treatment alone increased (P<0.001) mean plasma LH concentrations at day 26 but not at days 12 and 16 of lactation. In contrast, naloxone challenges given in the presence of Cabergoline treatment increased (P<0.05) mean LH at days 12 and 19 of lactation but not at day 26. Morphine challenges in the presence of Cabergoline treatment decreased (P<0.05) mean LH concentrations only at day 26 of lactation, but did not completely reverse the effect of Cabergoline. No treatment differences in plasma oestradiol-17β were detected at any time. Plasma prolactin decreased (P<0.001) in response to treatment with Cabergoline but there were no additional effects of naloxone or morphine. These data provide evidence for the existence of dopaminergic and opioidergic regulation of LH secretion in lactation in the sow and the relative influence of these systems changes as lactation progresses. Furthermore, the data suggest that the stimulatory effect of Cabergoline treatment on LH secretion in late lactation may be mediated by its effects on an inhibitory opioidergic mechanism. Finally, the data provide conclusive proof that prolactin does not directly influence LH secretion or estrogenic activity of the ovary during lactation in the sow.     

Changes in plasma progesterone and prolactin concentrations during the annual cycle and the role of prolactin in the maintenance of lactation and luteal development in the Antarctic fur seal (Arctocephalus gazella)

Progesterone in Antarctic fur seals was undetectable from 1-2 days before parturition to 4-6 days after parturition. There was a rapid increase in progesterone to 20 ng/ml between 6 and 10 days post partum and this increase coincided with peak concentrations of oestradiol-17 beta at the time normally associated with oestrus and mating in this species. Newly formed corpora lutea were present in the ovaries by Day 9 post partum even though the seals had been isolated in an enclosure and not mated. Thereafter, progesterone remained detectable, but at a low concentration (5 ng/ml) throughout embryonic diapause. A similar pattern was observed in unmated females which suggests they enter a period of pseudopregnancy. Progesterone increased to 35 ng/ml between late February and mid-March, indicating activation of the corpus luteum at the end of diapause, and then declined slowly through the remainder of gestation. Plasma prolactin, measured against a human prolactin standard, was elevated from 1-2 days before parturition and peaked at 0-3 days post partum. It then declined slowly throughout the post-partum period and remained at a low level throughout embryonic diapause. Prolactin concentration declined to undetectable at the end of diapause and before the end of lactation. Reduction of prolactin secretion by injections of bromocriptine from Days 3 to 5 post-partum terminated lactation. Mothers, which normally leave their pups to feed at sea on about Day 7 post partum, did not continue to lactate beyond Day 7 although this did not appear to be associated with reduced prolactin secretion. Bromocriptine treatment appeared to prevent the post-ovulatory surge of progesterone although there was no long-term effect of bromocriptine on progesterone secretion during the early stages of embryonic diapause/pseudopregnancy. This study has shown that prolactin is an important hormone for maintaining early lactation in the fur seal and it probably also has a role in the control of ovulation and luteal development. Prolactin does not appear to be implicated in the control of lactation cycles in fur seals. Changes in plasma progesterone during the annual cycle show that the pattern in fur seals resembles that of some carnivores with embryonic diapause.     

Ultrasonic cries from infant rats stimulate prolactin release in lactating mothers

Lactating female rats, separated from their litters for 6 hr, showed a dramatic elevation in plasma prolactin following a 15-min playback of ultrasonic vocalizations recorded from 7-day-old pups. Virgin females exhibited a smaller but still significant prolactin response to recorded pup calls. Control recordings of either background noise or adult ultrasonic vocalizations (22–26 kHz) had no effect on prolactin secretion even in lactating females. These results demonstrate that infant vocalizations can act as a stimulus for prolactin release and suggest that such communications may play a role in the maintenance of normal lactation.     

Effect of dopamine agonists or antagonists, TRH, stress and piglet removal on plasma prolactin concentrations in lactating gilts

The effect of dopamine agonists (ergocryptine), antagonists (chlorpromazine, haloperidol, reserpine, pimozide), thyrotropin releasing hormone or stress (restraint, piglet removal) on prolactin release was studied in primiparous lactating gilts. All animals were fitted with surgically implanted jugular catheters before farrowing. The only drug treatments which resulted in a significant change in PRL concentrations in blood were thyrotropin releasing hormone (increase) and ergocryptine (decrease). The results suggest that dopamine may not be the only regulator of prolactin in lactating pigs. Further studies are needed to identify drugs which would be useful in clinical situations for treatment of lactation failure due to low prolactin secretion. In the two stress-exposed groups, there was a gradual, steady decline in the plasma concentration of prolactin which resulted from loss of suckling contact with the piglets. Thus, snare restraint does not increase prolactin secretion in lactating sows confirming the results of other studies on pigs in different physiologic states.     

Prolactin does not mediate the suppressive effect of the suckling stimulus on luteinizing hormone secretion in ovariectomized lactating rats

The plasma LH concentration in ovariectomized lactating rats is low for 14 days postpartum, while the prolactin concentration is high during this period. We examined the effect of the inhibition of increased prolactin secretion with bromocriptine (CB-154) on the LH secretion in lactating rats ovariectomized on day 2 (day 0 = day of parturition). Blood samples were collected through an indwelling atrial cannula every day. LH levels were kept low until day 9 in lactating rats injected daily with CB-154 (0.6 mg/day, s.c.). The duration of the period during which LH secretion was suppressed was shorter in lactating rats treated with CB-154 than in saline-injected controls. The replacement with ovine prolactin by means of a mini-osmotic pump (0.3 mg/day, s.c.) in CB-154-treated lactating rats restored the duration of LH suppression. In rats deprived of their pups on day 2, the LH concentration rose immediately after removal of the pups and the LH level was not significantly different between rats treated with CB-154, ovine prolactin and saline, indicating that neither the CB-154 treatment nor the high level of prolactin alone has any effect on LH secretion in rats deprived of their pups. The present results clearly demonstrate that prolactin does not mediate the suppressing effect of the suckling stimulus on LH secretion in early lactation and support our theory that the suckling stimulus controls the LH and prolactin secretion independently at the hypothalamic level.     

Prolactin inhibition in lactating rats changes leptin transfer through the milk.

Malnutrition during lactation reduces milk production and changes pup's leptin serum levels. To test prolactin role in this nutritional state, we evaluated whether prolactin suppression during lactation changes serum leptin in dams, its transfer through the milk, and pup's serum leptin. Lactating rats were treated with bromocryptine (1 mg/twice a day, s.c.) or saline three days before sacrifice (days 2-4 or days 19-21). Food intake and body weight were measured until sacrifice (4th and 21st day). Serum prolactin and leptin were determined by radioimmunoassay. Bromocryptine injected dams had lower serum prolactin and milk production as expected. The mothers presented lower food ingestion (day 21: -25%), lower body weight (day 4: -12%; day 21: -10%), higher serum leptin (day 4: +68%), lower milk leptin on the 4th day (11 times) and higher (8 times) on the 21st day. The offspring of bromocryptine-treated mothers presented lower body weight in both periods of lactation and lower serum leptin on the 4th day (-40%) and higher on the 21st day (+37%) of lactation. We suggest that prolactin, through its effect on leptin secretion into the milk, may play an important role in signalizing maternal nutritional status to the pups.     

Evidence that suckling pups, through an exteroceptive mechanism, inhibit the milk stimulatory effects of prolactin in the rat during late lactation

Lactating rats were removed from the animal room on postpartum Day 20 and placed in a room where there were no other rats. On postpartum Day 21, these rats released prolactin either in response to exteroceptive signals from their own pups which were placed underneath or in response to those from 25 to 30 other lactating rats which along with their litters were placed in a rack 3 ft in front of the mothers' cage. Milk secretion was stimulated in the isolated rats on Day 21 by the prolactin released in response to exposure to the rack of lactators, but paradoxically was not stimulated by that released in response to exposure of the mother to her own pups. In fact, the stimulatory effects upon milk secretion, resulting from exposure to the rack of lactators was totally prevented if the mother was exposed to her pups 3–4 min before exposure to the rack of lactators. The blocking effect of the pups, however, did not occur when the pups were placed in cages alongside the mothers' cage. From subsequent experiments, it was concluded that sensory cues, from the pups, appeared to activate the sympathetico-adrenal system of the mother to release catecholamines which then blocked the milk stimulatory effects of prolactin which already had been discharged into the circulation. The pups did not inhibit milk secretion on Day 14 which indicates that the inhibiting mechanism becomes established sometime between Days 14 and 21 of lactation. These data suggest that a peripheral mechanism may operate to reduce milk secretion in the rat during late lactation, and thus, may be involved in the normal weaning process.     

Neuromedin U suppresses prolactin secretion via dopamine neurons of the arcuate nucleus

Neuromedin U (NMU) has a precursor that contains one additional peptide consisting of 33 or 36 amino acid residues. Recently, we identified this second peptide from rat brain and designated it neuromedin U precursor-related peptide (NURP), showing it to stimulate prolactin release from the pituitary when injected via the intracerebroventricular (icv) route. Here, we examined whether NMU, like NURP, also stimulates prolactin release. Unlike NURP, icv injection of NMU significantly decreased the secretion of prolactin from the pituitary. This suppression of prolactin release by NMU was observed in hyper-prolactin states such as lactation, stress, pseudopregnancy, domperidone (dopamine antagonist) administration, and icv injection of NURP. Immunohistochemical analysis revealed that icv injection of NMU induced cFos expression in dopaminergic neurons of the arcuate nucleus, but not the substantia nigra. Mice with double knockout of NMU and neuromedin S (NMS), the latter also binding to NMU receptors, showed a significant increase of the plasma prolactin level after domperidone treatment relative to wild-type mice. These results suggest that NMU and NURP may play important reciprocal roles in physiological prolactin secretion.     

Altered bile acid physiology during lactation in the rat

There is evidence that the rate of bile acid secretion increases significantly during lactation in the rat. We show that this increase in secretion rate is accompanied by an expanded bile acid pool and that occasioning the enhancement of both pool size and secretion is an increase in bile acid synthesis. The hypothesis is advanced that maternal prolactin, promoted by suckling young, amplifies cholesterol-7-alpha-hydroxylase, the rate-limiting enzyme in bile acid biosynthesis, and perhaps HMG-CoA reductase, the rate-limiting enzyme in cholesterogenesis.     

舒芬太尼联合地佐辛用于剖宫产术后镇痛的疗效及对患者血清泌乳素水平的影响

目的:探讨舒芬太尼联合地佐辛对剖宫产患者术后镇痛的效果及对患者血清泌乳素水平的影响。方法:选取我院收治的剖腹产患者90例,随机分为对照组及实验组,对照组给予舒芬太尼联合托烷司琼术后镇痛,实验组给予舒芬太尼联合地佐辛术后镇痛。观察并比较两组患者的镇痛效果及血清泌乳素PRL水平的影响,以及不良反应的发生情况等。结果:与术前比较,两组患者治疗后血清泌乳素PRL水平均升高,差异具有统计学意义(P0.05);与对照组比较,实验组治疗后血清PRL水平较高,差异具有统计学意义(P0.05);与对照组比较,实验组镇痛VAS评分、初次泌乳时间及泌乳充足时间、不良反应发生率均较低,差异具有统计学意义(P0.05)。结论:剖宫产术后应用舒芬太尼联合地佐辛的临床镇痛效果理想,在维持镇痛效果的同时,降低药物用量,减轻对脑垂体的刺激,提高泌乳素分泌,有效降低不良反应,值得临床推广应用。     

Regulation of ghrelin secretion during pregnancy and lactation in the rat: possible involvement of hypothalamus

We investigated the plasma concentration of ghrelin peptide during pregnancy and lactation in rats. Plasma ghrelin levels on days 10 and 15 of pregnancy were significantly lower than those of the non-pregnant rats. Thereafter, the plasma ghrelin levels on day 20 of pregnancy sharply increased to levels comparable with those in non-pregnant rats. Ghrelin peptide concentrations in the stomach did not change significantly during pregnancy. In the hypothalamus, ghrelin mRNA levels were significantly lower on day 15 of pregnancy than in the non-pregnant rats. Also, plasma ghrelin levels were significantly lower in lactating dams than non-lactating controls on days 3 and 8 of lactation. We examined the possible involvement of prolactin and oxytocin in the regulation of plasma ghrelin concentrations during lactation. Although plasma prolactin levels were decreased by the administration of bromocriptine, plasma ghrelin levels did not differ significantly between vehicle- and drug-treated lactating rats. Administration of haloperidol produced a marked increase in plasma prolactin levels as compared with the non-lactating controls. However, plasma ghrelin levels were not significantly different between vehicle- and drug-treated rats. Administration of an oxytocin antagonist into the lateral ventricle significantly inhibited the increase in the plasma oxytocin level induced by acute suckling. However, plasma ghrelin levels did not significantly between the groups. These observations indicated that the decrease in serum ghrelin is caused by a loss of the contribution of hypothalamic ghrelin. Furthermore, the present results suggested that the suckling stimulus itself, but the release of prolactin or oxytocin, is the factor most likely to be responsible for the suppression of ghrelin secretion during lactation.