Prolactin Receptor mRNA Expression Research Articles

Regulatory effects of prolactin in amphibians

Prolactin (PRL) is a protein hormone of the anterior pituitary and has a diverse role in different aspects of amphibian life - in courtship, reproduction, larval development and in other physiological functions. The expression of PRL receptor mRNA is reported in skin, gill, liver, gut, kidney, gonads and many other organs of different groups of amphibians. PRL stimulates growth and inhibits the metamorphosis of larvae of several species of amphibians. PRL causes seasonal changes in sex accessory structures and secondary sexual characteristics. PRL affects metabolic regulation and suggests phylogenic conservation of its receptors. Plasma PRL concentration changes during the annual reproductive cycle in many amphibians. In courtship behavior of newt, PRL also induces the synthesis of the newt courtship pheromones. PRL acts on skin and its function is also associated with water drive in amphibians. PRL causes water retention in amphibians. PRL acts centrally or peripherally to induce the courtship behavior. The key goal of the paper is to understand the roles of PRL in various aspect of amphibian life, like in aspects of morphology, behaviour, physiological functions and how PRL interfere amphibian life in presence and absence of other chemical and biological factors.

The role of prolactin in the suppression of the response to restraint stress in the lactating mouse.

Suppression of the hypothalamic-pituitary-adrenal (HPA) axis is a well-characterised maternal adaptation that limits the exposure of the offspring to maternally-derived stress hormones. This current study has investigated the possible involvement of the lactogenic hormone, prolactin, in this physiologically important adaptation. As expected, circulating prolactin levels were higher in unstressed lactating mice compared to their virgin counterparts. Interestingly however, the ability of an acute period of restraint stress to further elevate prolactin levels was diminished in the former group. The stress-induced rise in prolactin levels in the virgin animals was concurrent with an increase in prolactin receptor activation within the adrenal cortical cells. This adrenal response was not seen in either the stressed or control lactation group, an observation that may be in part explained by the observed downregulation of prolactin receptor mRNA expression within this tissue. Further evidence of suppression of the HPA axis during lactation was revealed using in situ hybridisation to demonstrate that while acute restraint stress increased corticotrophin releasing hormone (CRH) mRNA expression in the hypothalamic paraventricular nucleus in both virgin and lactating mice, the magnitude of this response was reduced in the latter group. This potentially adaptive response did not, however, appear to result from the altered prolactin profile during lactation because it was not affected by the pharmacological suppression of prolactin secretion from the pituitary. This study therefore suggests that during lactation the response of the HPA axis to stress is suppressed at multiple physiological levels which are mediated by both prolactin-dependent and prolactin-independent mechanisms.

Postpartum Fatigue and Inhibited Lactation

Background:Postpartum fatigue is a common disorder worldwide and affects both physical and mental functioning. In breastfeeding women, Prolactin (PRL) is not only involved in immunoregulation, but also responsible for lactation. Prolactin levels in women with chronic fatigue are higher than normal, but a chronic fatigue state inhibits postpartum lactation in humans. Objectives:This paper explored the inhibition mechanism of lactation by postpartum fatigue in rats.Methods:Postpartum fatigue models were built by forcing mother rats to stand in water and divided into 3-hour, 9-hour and 15-hour per day fatigue groups according to the underwater time. Mother rats and their offspring were reunited in a dry cage for 90 minutes every 3 hours for feeding. The expression of PRL, PRL receptor (PRLR), Janus Kinase 2 (JAK 2), and Signal transducers and activators of transcription 5 (STAT5) mRNA were analyzed and the microstructure of mammary gland were observed under light and electron microscopy. Results:The expression of pituitary PRL mRNA and its downstream signaling pathway JAK2 and STAT5 mRNA were down-regulated in the severe postpartum fatigue rats. PRL mRNA responses were dose-related to duration of fatigue. The expression of PRLR mRNA increased. Postpartum fatigue led to functional degeneration of mammary gland. The breast lobules were shrunk and the number of alveoli were decreased. Few milk protein granules and fat droplets were observed in the cytoplasm under transmission electron microscope. Conclusion:Postpartum fatigue inhibits the lactation by down-regulating the expression of PRL and PRL-dependent signaling pathway in rats.

Pregnancy-induced changes in the transcript levels of prolactin receptor and its suppressor in the ovine hypothalamus and adenohypophysis.

The aim of this study was to analyse changes in the abundance of prolactin (PRL) receptor (PRLR) and suppressor of cytokine signalling-3 (SOCS-3) mRNA in the ventro-/dorsomedial nucleus (VMH/DMH) and arcuate nucleus (ARC) of the hypothalamus as well as in the median eminence (ME) and adenohypophysis (AP) in sheep at 30, 60, 90 and 120 d of pregnancy compared to non-pregnant animals. In the VMH/DMH, PRLR transcripts were detected only in non-pregnant ewes. In the ARC, the abundances of PRLR mRNA were higher in pregnant sheep on days 30 (p<.01), 90 (p<.01) and 120 (p<.05) than in non-pregnant sheep. In contrast, the expression of PRLR mRNA in the ME was lower (p<.01) in pregnant ewes at days 30 and 60 than in non-pregnant ewes and was undetectable at later stages of gestation. In all studied stages of pregnancy except day 60, the abundance of PRLR mRNA was higher (p<.01) in the ARC than in the AP, while in non-pregnant sheep, there were no differences (p≥.05) in the transcript levels between these two tissues. In non-pregnant ewes, the abundance of SOCS-3 mRNA in the AP was lower than that in any other studied tissue (p<.05-p<.01). In conclusion, the observed changes in PRLR and SOCS-3 mRNA abundance in the hypothalamus and AP during pregnancy may be important components of the mechanisms regulating the action of PRL in energy homeostasis and neuroendocrine interactions within the hypothalamic-pituitary axis.

Reproductive status affects the expression of prolactin receptor mRNA in the brain of female Damaraland mole-rats

The eusocial Damaraland mole-rat (Fukomys damarensis) represents an extreme example of reproductive skew, in that reproduction is completely blocked in female subordinate group members. It is thought that in these animals normal GnRH secretion from the hypothalamus is disrupted. Prolactin, a peptide hormone secreted from the anterior pituitary gland, has been implicated in a wide variety of functions. Well documented in rodents is its role in mediating lactational infertility. Elevated circulating prolactin levels, such as during lactation, are associated with reduced GnRH release into the portal blood and with a reduction in the frequency and amplitude of LH pulses. The present study aimed at investigating whether such a mechanism could act in reproductively suppressed female Damaraland mole-rats. By means of in situ hybridisation we studied the distribution and gene expression of the prolactin receptor (Prlr) in wild-caught female Damaraland mole-rats with different reproductive status. Substantial Prlr expression was found in several brain regions, with highest levels in the choroid plexus and moderate expression in the preoptic and tuberal hypothalamus. While in reproductive and non-reproductive females plasma prolactin levels were very low and not significantly different, quantification of the Prlr hybridisation signal revealed significant differences in relation to reproductive status. Reproductively suppressed females had increased expression of Prlr in the choroid plexus and in the arcuate nucleus (ARC) when compared to reproductive females. This suggests higher local prolactin levels in the brain of suppressed females. Together with previous findings, it could indicate that prolactin inhibits ARC kisspeptin neurons, which then would lead to reduced activation of GnRH neurons in such females.

Expression of Prolactin Receptor mRNA in Lactotrophs and Somatotrophs of the Chicken Anterior Pituitary Gland.

Prolactin (PRL) is a hormone mainly secreted by the anterior pituitary gland. In birds, PRL exerts a variety of physiological functions in target tissues expressing the PRL receptor (PRLR). In chicken, the PRLR mRNA is abundant in the anterior pituitary gland, but its regional and cellular localization are unknown. In the present study, we investigated the expression of the PRLR mRNA in cephalic and caudal lobes of the chicken anterior pituitary gland. Real-time polymerase chain reaction (PCR) revealed high levels of PRLR mRNA in both cephalic and caudal lobes. In situ hybridization revealed that the PRLR mRNA was distributed in a wide area of both lobes, and co-localized with the PRL and growth hormone (GH) mRNAs in the cephalic and caudal lobes, respectively. These results suggest that PRL exerts autocrine/paracrine effects through PRLR on PRL-producing lactotrophs and GH-producing somatotrophs in the chicken anterior pituitary gland.

Essential role of endogenous prolactin and CDK7 in estrogen-induced upregulation of the prolactin receptor in breast cancer cells.

Our early studies have shown that Estradiol (E2)/Estrogen Receptor α (ER) in a non-DNA dependent manner through complex formation with C/EBPβ/SP1 induced transcriptional activation of the generic hPIII promoter and expression of the Prolactin Receptor (PRLR) receptor in MCF-7 cells. Subsequent studies demonstrated effects of unliganded ERα with requisite participation of endogenous PRL on the activation of PRLR transcription. Also, EGF/ERBB1 in the absence of PRL and E2 effectively induced upregulation of the PRLR. In this study we have delineated the transcriptional mechanism of upregulation of PRLR receptor induced by E2 incorporating knowledge of the various transcriptional upregulation modalities from our previous studies. Here, we demonstrate an essential requirement of STAT5a induced by PRL via PRLR receptor which associates at the promoter and its interaction with phoshoERα S118. Knock-down of PRL by siRNA significantly reduced E2-induced PRLR promoter activity, mRNA and protein expression, recruitment of ERα to the complex at promoter, C/EBPβ association to its DNA site and productive complex formation at hPIII promoter. The specific CDK7 inhibitor (THZ1) that attenuates E2-induced ERα phosphorylation at S118 abrogated E2-induced PRLR promoter activation. Further studies demonstrated that E2 induced cell migration was inhibited by PRL siRNA and THZ1 indicating its dependence on PRL/PRLR and CDK7, respectively. Our studies have demonstrated the essential role of endogenous PRL and CDK7 in the upregulation of PRLR by E2 and provide insights for therapeutic approaches that will mitigate the transcription/expression of PRLR and its participation in breast cancer progression fueled by E2 and PRL via their cognate receptors.

Evidence of a role for prolactin as regulators of ovarian follicular development in goose

Background: Prolactin (PRL) regulates development and reproduction, and its effects are mediated by the prolactin receptor (PRLR). In order to clarify the role of PRLR and PRL in the process of follicular development in the goose ovary, the level of PRLR mRNA expression in the ovary and follicles of the Sichuan white goose was determined, as well as the PRL concentration in ovarian follicles. Results: The level of PRLR mRNA in the hierarchical follicles (HFs) initially increased, and subsequently decreased, whereas PRLR expression was initially low and later increased in postovulatory follicles (POFs). The level of PRLR mRNA expression was the highest in the F4 follicles, and lowest in the F1 follicles in all of the examined follicles. Compared with the level of PRLR mRNA expression in the small white follicles (SWFs), the level of PRLR mRNA was 2.86- and 1.44-fold higher in the F4 and small yellow follicles (SYFs), respectively (P Conclusions: The change of PRL concentration was similar to the PRLR mRNA expression level in preovulatory follicles. These results suggest that the PRL mediated by the PRLR plays a stimulatory role in the SWF to SYF transition.

MRNA expression of prolactin receptor in sheep pituitary and its effect on maternal behavior.

Prolactin receptor (PRLR) is the specific receptor for prolactin. Brain-specific control of PRLR gene expression is correlated with the induction of maternal behavior. The aim of this study was to investigate the dynamic changes in the PRLR mRNA level in the sheep pituitary gland and its effect on ewe maternal behavior. The mRNA expression of PRLR during non-pregnancy stage was maintained at low but detectable levels, as was the case during mid-gestation. After that, expression began increasing and reached a maximum during the early postpartum followed by a gradual decline during the mid-postpartum and late postpartum stages. A trend of higher PRLR mRNA expression was observed during the postpartum stage relative to the non-pregnancy and pregnancy stages. Expression of PRLR mRNA was significantly higher in the normal maternal behavior group than that in the abnormal maternal behavior group. There may be a correlation between PRLR mRNA expression and ewe maternal behavior, especially in early postpartum.

Identification of bovine prolactin in seminal fluid, and expression and localization of the prolactin receptor and prolactin-inducible protein in the testis and epididymis of bulls exposed to ergot alkaloids

The objectives of this study were to determine (1) the presence and expression levels of bovine prolactin receptor (PRLR) and prolactin-inducible protein (PIP) in bovine testis and epididymis, and (2) the presence and concentrations of prolactin (PRL) present in seminiferous fluid in bulls consuming diets with (E+) or without (E−) ergot alkaloids. Bulls (n = 8) were sacrificed after 126 days (group A) of E+ or E− treatment or 60 days after all bulls (n = 6) were switched to the E− ration (group B). End point and real-time quantitative reverse transcription–polymerase chain reaction and immunohistochemistry were conducted on testis and epididymis samples to establish the presence and relative expression of PRLR and PIP. Seminal fluid samples obtained from bulls consuming E− and E+ diets were subjected to RIA for PRL. Both PIP and PRLR were present in testis and epididymis as determined by reverse transcription–polymerase chain reaction and immunohistochemistry. Prolactin-inducible protein mRNA abundance was affected by time of slaughter in testis and epididymis head, respectively (P < 0.05). Prolactin receptor mRNA expression was affected by time of slaughter in the epididymis (P < 0.05) and differed in testis samples because of treatment (P < 0.05). Radioimmunoassay establishes the presence of PRL in seminal fluid; however, differences in the concentration of PRL over two separate studies were inconsistent, possibly because of differences in diet. The presence and localization of the PRLR are consistent with expression data reported for other species, and the presence of PIP and PRL in seminal fluid is consistent with data generated in humans.

AB0033 Systemic secreted prolactin in ra leads to proinflammatory cytokine production in macrophages

BackgroundRheumatoid arthritis (RA) is a rheumatic disease mainly affecting women. It is known that prolactin (PRL) is a sex hormone, which apart from inducing lactation, also has immunomodulatory properties. PRL...

In vitro Suppression of Prolactin During Later Stages of Egg Lay in Domestic Hen (Gallus gallus Domesticus) Anterior Pituitcytes by RNA Interference

2 Abstract: Prolactin (PRL) is a peptide hormone secreted by anterior pituitary gland. PRL increases as per the age and reproductive cycle of birds from normal physiological levels to extremely higher level there by affecting the ovulation, egg formation, oviposition, egg production and hyperprolactinemia. This is mor e pronounced and persistent after 72 weeks of age in birds. Hence a study was conducted in anterior pituitary primary cultured cells obtained from 72 and 82 weeks old white leg horn (WLH) hens to knock down the PRL gene expression by siRNA and observing its effects on PRL, PRL mRNA, protein content of PRL, prolactin receptor (PRLR) and Growth Hormone (GH) mRNA to unravel the functional role of PRL at 72 and 82 week age by siRNA for PRL. Three siRNAs were designed as per standard siRNA protocols and studied the suppression of PRL gene expression in primary cultured cells procured from adult chicken anterior pituitary glands in in vitro conditions. Average percentage reduction of PRL in anterior pituitary primary cell culture following siRNA transfection was 82 and 60% at 72 and 82 weeks respectively. Protein content of PRL was significantly (p<0.01) decreased in siRNA transfected cells compared to controls. Growth Hormone (GH) mRNA and PRL receptor (PRLR) mRNA levels did not change significantly (p<0.01) between control and treated cells. Results clearly suggested that the siRNA designed for PRL specifically decreased PRL gene expression in in vitro conditions. Level of PRLR mRNA and GH mRNA levels expression did not follow the similar pattern of PRL gene expression in anterior pituitary cells. It is concluded that, construction of short specific siRNA for PRL significantly decreased PRL, PRL mRNA and protein content of PRL without showing any effect on PRLR and GH between the two age groups of birds. These results may lead to construction of short specific siRNA for stable and chronic suppression of PRL gene expression during embryogenesis before an increase of PRL occurs for long term knock down of PRL in in vivo conditions. In conclusion , understanding of hyperprolactinemia and the involvement of PRL may provide the basis for the development of therapeutic drugs or methods against hyperprolactinemia by RNA interference.

Regulation of Prolactin Receptor Gene Expression in the Rat Choroid Plexus via Transcriptional Activation of Multiple First Exons during Postnatal Development and Lactation

Prolactin (PRL) has numerous physiological functions that are mediated by its receptors in target cells. Expression of the rat PRL receptor (PRLR) gene is regulated in a tissue-specific manner via the transcriptional activation of five distinct first exons, i.e., E1(1), E1(2), E1(3), E1(4), and E1(5). In the present study, we investigated the expression profiles of these first exon variants of PRLR mRNA in the rat choroid plexus, which is considered to be a site of receptor-mediated PRL transport from the blood to cerebrospinal fluid. Real-time PCR analysis revealed that E1(3)-, E1(4)-, and E1(5)-PRLR mRNA expression levels increased in the choroid plexus in male and female rats during postnatal development, with markedly higher level of E1(4)-PRLR mRNA. In female rats, the E1(4)-PRLR mRNA expression levels increased markedly during lactation compared with the diestrus state, whereas there was no increase in the E1(3)- and E1(5)-PRLR mRNA levels. The E1(4)-PRLR mRNA expression pattern was similar to that of the total PRLR mRNA. The PRL plasma concentration generally correlated with the E1(4)-PRLR mRNA expression levels in both sexes. These findings suggest that PRLR gene expression in the choroid plexus is upregulated mainly via the transcriptional activation of the E1(4)-first exon.

Physiological and hormonal aspects in female domestic pigeons (Columba livia) associated with breeding stage and experience

The present study examined the changes in serum biochemical values, hormone profiles and ovary prolactin receptor (PRLR) gene expression occurring in female domestic pigeons (Columba livia) under different breeding status and experience. The egg-laying pigeons had lower calcium, total protein, albumin, prolactin levels and higher oestrogen levels than those of incubating birds (p < 0.05). First-time breeders had higher (p < 0.05) progesterone levels and lower (p < 0.05) prolactin levels than that of experienced ones. The levels of oestrogen and follicle-stimulating hormone (FSH) increased with age (p < 0.05). The very old birds showed a pronounced increase (p < 0.05) in PRL, FSH and progesterone and a little decrease in oestrogen. Serum prolactin level was not correlated with the ovary PRLR mRNA expression pattern among all the pigeons. Results showed that serum physiological profile of female pigeons was correlated with breeding status, whereas reproductive hormone levels were correlated with advancing breeding experience. It was concluded that female pigeons had a good ability of recovering from nutrient loss after each breeding attempts, and the degradation of reproductive performance might be attributed to changes in the endocrine system.

Expression of prolactin receptors in normal canine mammary tissue, canine mammary adenomas and mammary adenocarcinomas

BackgroundMammary tumors represent the most common neoplastic disease in female dogs. Recently, the promoting role of prolactin (PRL) in the development of human breast carcinoma has been shown. Possible proliferative, anti-apoptotic, migratory and angiogenic effects of PRL on human mammary cancer cells in vitro and in vivo were suggested. The effects of PRL are mediated by its receptor, and alterations in receptor expression are likely to play a role in tumor development. Currently, not much data is available about prolactin receptor (PRLR) expression in canine mammary tumors. To set the basis for investigations on the role of PRL in mammary tumorigenesis in this species, prolactin receptor expression was evaluated by semi-quantitative real time PCR and immunohistochemistry on 10 formalin-fixed, paraffin-embedded samples each of canine non-neoplastic mammary tissue, mammary adenomas and adenocarcinomas.ResultsThe highest PRLR expression levels were found in normal mammary tissue, while adenomas, and to an even higher degree adenocarcinomas, showed a significant decrease in prolactin receptor expression. Compared to normal tissue, PRLR mRNA was reduced 2.4 fold (p = 0.0261) in adenomas and 4.8 fold (p = 0.008) in adenocarcinomas. PRLR mRNA expression was significantly lower in malignant than in benign lesions (p = 0.0165). Immunohistochemistry demonstrated PRLR expression in all three tissue types with signals mostly limited to epithelial cells.ConclusionsMalignant transformation of mammary tissue was associated with a decline in prolactin receptor expression. Further studies are warranted to address the functional significance of this finding.

The prolactin receptor mediates HOXA1-stimulated oncogenicity in mammary carcinoma cells

The HOX genes are a highly conserved subgroup of homeodomain-containing transcription factors that are crucial to normal development. Forced expression of HOXA1 results in oncogenic transformation of immortalized human mammary cells with aggressive tumour formation invivo. Microarray analysis identified that the prolactin receptor (PRLR) was significantly upregulated by forced expression of HOXA1 in mammary carcinoma cells. To determine prolactin (PRL) involvement in HOXA1‑induced oncogenicity in mammary carcinoma cells (MCF-7), we examined the effect of human prolactin (hPRL)-initiated PRLR signal transduction on changes in cellular behaviour mediated by HOXA1. Forced expression of HOXA1 in MCF-7 cells increased PRLR mRNA and protein expression. Forced expression of HOXA1 also enhanced hPRL-stimulated phosphorylation of both STAT5A/B and p44/42 MAPK, and increased subsequent transcriptional activity of STAT5A and STAT5B, and Elk-1 and Sap1a, respectively. Moreover, forced expression of HOXA1 in MCF-7 cells enhanced the hPRL‑stimulated increase in total cell number as a consequence of enhanced cell proliferation and cell survival, and also enhanced hPRL-stimulated anchorage-independent growth in soft agar. Increased anchorage-independent growth was attenuated by the PRLR antagonist ∆1-9-G129R‑hPRL. In conclusion, we have demonstrated that HOXA1 increases expression of the cell surface receptor PRLR and enhances PRLR-mediated signal transduction. Thus, the PRLR is one mediator of HOXA1‑stimulated oncogenicity in mammary carcinoma cells.

Molecular characterization, mRNA expression of prolactin receptor (PRLR) gene during pregnancy, nonpregnancy in the yak ( Bos grunniens)

Prolactin (PRL) plays central roles in a wide range of body functions in mammals, and the actions are mediated by the specific cell surface receptor, the prolactin receptor (PRLR). To better understand the role of PRL in the yak ( Bos grunniens), in the present study, we first cloned yak PRLR cDNA, and compared its mRNA expression in several tissues with cattle ( Bos taurus). By reverse transcriptase-polymerase chain reaction (RT-PCR) strategy, we obtained full-length of yak PRLR cDNA sequence comprised of an open reading frame of 1746 bp encoding a 581 amino acid protein, and contained a signal sequence and a transmembrane region. The intracellular domain had two pairs of cysteine residues and a WSXWS motif. The cytoplasmic domain comprised 323 residues and contained box 1 sequence. The yak PRLR shared 66.0–98.5% protein sequence identity with mammalian homologs. Real-time PCR analysis revealed that PRLR mRNA was higher in mammary tissue than in ovary and endometrium ( P < 0.01). During pregnancy, the ovary and mammary PRLR mRNA expression increased by 33- and 2.9-fold in yak, respectively, and increased by 46- and 3.8-fold in cattle, respectively. PRLR mRNA expression was higher ( P < 0.05) in mammary tissue and ovary of pregnant cow than that of pregnant yak. It is proposed that the increased ovarian and mammary PRLR mRNA expression during pregnancy may be associated with corpus luteum function for maintenance of pregnancy and mammary development for subsequent lactation.

Expression of Prolactin Receptor mRNA after Melatonin Manipulated in Cashmere Goats Skin during Cashmere Growth

The aim of this research was to investigate the dynamic changes of the level of total prolactin receptor (PRLR) mRNA and the short form prolactin receptor (S-PRLR) mRNA in skin of cashmere goats from the initiation of cashmere fibre growth to active growth. Eighteen half-sib wethers were allocated randomly to two groups. Melatonin implants were used in order to initiate growth of cashmere fibre before the normal time and reduce blood plasma prolactin (PRL) concentration. Real-time reverse transcription quantitative polymerase chain reaction (real-time PCR) was used to determine PRLR mRNA expression levels of skin from June to November. The results showed that, in Chinese Inner Mongolia cashmere goats, there were seasonal variations in expression of total PRLR mRNA in skin with levels decreasing from June to October. Synchronously, the cashmere fibre growth rate gradually increased during this period, but the expression levels of S-PRLR mRNA did not decrease along with seasonal variation from initiation to active growth of cashmere fibre. These results suggest that expression levels of S- PRLR mRNA might be involved in the process of cashmere growth. It was also possible that the change of alternative splicing of PRLR occurred in the skin of cashmere goats from proanagen to anagen.

Prolactin induces phosphorylation of the STAT5 in adrenal glands of Hatano rats during stress

Aims To investigate the signaling of prolactin (PRL) in the adrenal gland during stress in Hatano high- (HAA) and low-avoidance (LAA) rats. Main methods Adrenal glands of both strains were collected at 0, 15 and 30 min after stress. The protein levels of phosphorylated STAT5 and the mRNA levels of melanocortin receptor 2 (MC2R) and PRL receptor (PRLR) were analyzed. Furthermore, the effects of bromocriptine-induced hypoprolactinemia on adrenocortical responses to stress were investigated. Key findings Adrenocorticotropic hormone (ACTH) concentrations in HAA were greater than LAA, while the difference in PRL concentrations were found only at 120 min after stress induction. No strain differences were observed in corticosterone or progesterone in response to stress. The stress-induced increase in MC2R mRNA expression was higher in HAA, but there was a lowered PRLR mRNA expression. STAT5 become highly phosphorylated in response to stress in both strains, but bromocriptine led to a reduction the STAT5 phosphorylation. Exposure to bromocriptine was associated with a reduction in plasma PRL in response to stress in both strains, while the ACTH levels were not altered. However, the decrease in corticosterone and progesterone in response to stress was observed only in bromocriptine-treated LAA rats. Significance These data show that PRL plays an important role in the regulation of corticosterone and progesterone release in LAA but not in HAA during stress. These results suggest that PRL increase in response to stress, and it acts on the adrenal cortex and thereby plays an important physiologic role in protecting against acute stress.

249 THE DIRECT AND CUMULUS-MEDIATED EFFECTS OF PROLACTIN ON BOVINE OOCYTES

Prolactin (PRL) is an important regulator of female reproduction, as indicated using PRL receptor knockout mice (Bole-Feysot C et al. 1998 Endocrinol. Rev. 19, 225–268). The presence of PRL receptors or their mRNA has been shown in different follicular compartments of various mammalian species, including oocytes of sheep and mice (Picazo RA et al. 2004 Reproduction 128, 545–553; Kiapekou E et al. 2005 Reprod. Biomed. Online 10, 339–346). The aim of the present study was to characterize the direct and cumulus-mediated pathways of PRL signaling into bovine oocytes. The expression of PRL receptor mRNA in follicular cells was detected by reverse transcription polymerase chain reaction (RT-PCR) analysis. For oocytes, nested PCR was used. In addition, a total of 707 COC and 689 denuded oocytes (DO) from follicles 2 to 8 mm in diameter were cultured for 6 h (COC), 8 h (DO), or 24 h (COC and DO) in the presence or absence of 50 ng mL–1 bovine PRL (20 IU mg–1; Research Center for Endocrinology, Moscow, Russia). The following systems for COC and DO culture were used: (1) TCM-199 containing 10% fetal calf serum (system 1) and (2) DMEM containing 10 U mL–1 pregnant mare’s serum gonadotropin (PMSG), 5 U mL–1 hCG (Intervet Scandinavia, Copenhagen, Denmark), and 5% estrous cow serum (system 2). The nuclear status of oocytes was evaluated by Tarkowski’s cytogenetic method. After IVM in system 2, a portion of the oocytes (348 COC and 311 DO) underwent IVF and IVC, and the embryo development was tracked until the blastocyst stage. All treatments were repeated 4 to 7 times. Results were expressed as mean ± SEM. Arcsine-transformed data were analyzed by two-way ANOVA. Messenger RNA expression of long and short isoforms of PRL receptor was revealed in both bovine oocytes and cumulus cells. In system 1, PRL raised the rate of DO remaining at the diplotene stage by 8 h of culture (from 32.6 ± 3.0 to 47.5 ± 4.7%, P &lt; 0.05), whereas there was no effect of PRL on the meiotic resumption in COC. By contrast, the hormone exerted a stimulatory action on the meiotic progression in the presence of cumulus cells by increasing the proportion of oocytes reaching the telophase I or metaphase II stages during 24 h of maturation (from 61.9 ± 1.2 to 73.5 ± 0.9%, P &lt; 0.05). In system 2, PRL did not affect nuclear maturation of either cumulus-enclosed or cumulus-free oocytes, with the maturation rate varying between 93.9 ± 2.7 and 95.2 ± 2.4% (COC) and between 85.6 ± 3.0 and 83.0 ± 4.0% (DO). When added to IVM system 2, PRL raised the cleavage and blastocyst rates only in the case of COC (from 72.2 ± 2.4 to 80.2 ± 2.1%, P &lt; 0.05, and from 20.5 ± 3.6 to 40.9 ± 4.2%, P &lt; 0.01, respectively). These findings suggest functioning of the direct and cumulus-mediated pathways of PRL signaling into bovine oocytes, with the hormone affecting meiosis via both pathways. We thank A. D. Roed for expert technical assistance with IVF and IVC. This research was supported in part by RFBR (project no. 07-04-01485). I.L. was the recipient of an Erasmus Mundus fellowship.