Non-genomic actions of progesterone and estrogens in regulating reproductive events in domestic animals

It has been established that nuclear receptors mediate the action of estrogens and progestins in regulating gene expression in the hypothalamic-hypophyseal-gonadal axis of domestic animals during various reproductive states. Results of recent in vitro studies suggest that estradiol-17beta and progesterone can act non-genomically to affect signal transduction responses in target cells by binding to receptors in the plasma membrane. The genomic action of steroids is generally detectable in hours to days whereas non-genomic responses of cells occur in seconds to minutes. The nature of the plasma membrane receptors for estrogens and progesterone has been explored but has not been conclusively established for all cell types studied. In the ewe, estradiol-17beta or estradiol-bovine serum albumin conjugate has been shown by in vitro and in vivo approaches to act non-genomically to suppress luteinizing hormone secretion by gonadotropes and stimulate production of nitric oxide by uterine arterial endothelial cells. Progesterone has been shown to inhibit oxytocin (OT) binding to its receptor in isolated ovine endometrial plasma membranes. This non-genomic action of progesterone blocks OT activation of the phosphoinositide cascade and production of prostaglandin F(2alpha) by ovine and bovine endometrium. The acrosome reaction of caprine and porcine spermatozoa is activated by the non-genomic action of progesterone. Further research is required to define the biological significances of the non-genomic actions of estrogens and progestins.     

The Role of Uterine and Ovarian Hormones in Luteolysis: A Comparison among Species

Luteolytic mechanisms have evolved in mammals to improve reproductive efficiency. The hormonal interactions that control the onset and progress of luteolysis are complex. They involve endocrine and paracrine signals that link the corpus luteum, uterus and posterior pituitary gland. Current concepts concerning these interactions will be examined in the five major domestic ungulate species commonly raised in Europe and North America (cattle, sheep, goats, pigs and horses). Some of these interactions are similar across species. All five depend on prostaglandin F_2α secreted from the uterus, to induce luteolysis. Three hormones, progesterone, estradiol and oxytocin interact to regulate uterine secretion of PGF_2α. Oxytocin is an acute stimulus for uterine PGF_2α secretion. Progesterone and estradiol interact to regulate uterine secretary responsiveness to oxytocin. Precisely how these hormones interact varies across species.     

Endometrial Oxytocin Receptor Concentration and Activity in Prepubertal Ewe Lambs

We have measured endometrial oxytocin receptor concentrations during prepuberty in ewe lambs, and have investigated the effect of progesterone on the activity of these receptors. In the first study, oxytocin receptor concentrations were undetectable in 2-week prenatal lambs but had increased immediately following birth and were then maintained throughout prepubertal life. Despite the presence of oxytocin receptors animals showed no prostaglandin F(2alpha) (PGF(2alpha)) release in response to exogenous oxytocin challenge at either 3 or 5 months of age. In a second study in 4-month-old ewe lambs treatment with exogenous progesterone resulted in the appearance of PGF(2alpha) release in response to oxytocin after 10 days of treatment. Thus, during the prepubertal life, ewe lambs possess the prerequisites of a luteolytic mechanism in that they have a dormant population of oxytocin receptors in which progesterone can induce oxytocin-stimulated PGF(2alpha) release.     

Steroids as local regulators of ovarian activity in domestic animals

The presented overview gives clear evidence for steroids as local regulators of follicular and luteal activity. In the follicle, estrogen receptor-alpha (ERalpha) and ERbeta expression are demonstrated in cow, ewe and pig. Besides species specific effects in general, there is evidence that estradiol-17beta (E(2)) exerts a dose-dependent inhibition on the secretion of progesterone (P(4)) by both theca interna cells (TI) and granulosa cells (GC). GC enhance the ability of the TI to produce androstendione by supplying them with progestin precursor. Androgen produced by TI enhances the ability of the GC to make E(2), and high concentrations of E(2) in the preovulatory follicle inhibit 3beta-HSD in both TI and GC and thus, may promote the use of the pathway Delta(5) for TI androgen production. The authors suggest that E(2) acts within the follicle to exert positive feedback on androgen and E(2) production, and exerts mitotic and anti-atretic or anti-apoptotic effects on follicular cells. Parts of the E(2)-mediated local action are regulated by stimulating effects on hormone receptors (LH, FSH, oxytocin). Gap junctions permit transfer of nutrients and cytokines to and from the avascular GC and oocyte, and formation is stimulated by estrogens. In bovine corpus luteum (CL) there is evidence that P(4) may directly regulate the production of P(4), oxytocin and prostaglandins (PGs) in a cycle dependent fashion. In most of domestic animal species, there is clear evidence for CL production of E(2) with clear stimulatory and luteotropic effects on P(4), and an intraluteal circuit that involves paracrine effects of E(2), oxytocin and PGF(2alpha) (especially in pigs). In contrast, there are species (ruminants, mares) in which the evidence for important local effects of E(2) is less clear, although expression of ERalpha, ERbeta and progesterone receptor (PR) is documented. Progesterone is very important for the regulation of CL lifetime by effects on the endometrium and release of the luteolytic signal PGF(2alpha). In conclusion, steroids as local regulators of ovarian activity are now documented and may stimulate further research in this field.     

Hormonal regulation of oxytocin-induced prostaglandin F(2alpha) secretion by the bovine and ovine uterus in vivo

In long-term ovariectomized ewes and cows, endometrial oxytocin receptors rest at relatively high levels but oxytocin is unable to induce prostaglandin F(2alpha) release. A series of studies were carried out to investigate the roles of physiological levels of progesterone and estradiol in "activating" these receptors in terms of permitting oxytocin-induced prostaglandin F(2alpha) release. In long-term ovariectomized cows, treatment with progesterone, but not estradiol, resulted in the induction of responsiveness to oxytocin. This responsiveness appeared within 2 d of progesterone treatment, reached a maximum by 6 d and was maintained to Day 18. In ovariectomized ewes, while estradiol treatment did induce temporary responsiveness to oxytocin after 3 d of treatment, treatment with progesterone was required to induce sustained responsiveness that appeared by Day 9 of treatment and was maintained to Day 12. Measurement of endometrial receptors for oxytocin revealed a significant decline in oxytocin receptors by Day 6 of progesterone treatment when responsiveness to oxytocin was maximal, demonstrating that receptor concentrations were not a limiting factor. The most likely mechanism by which progesterone treatment induces responsiveness to oxytocin may be through the up regulation of post receptor signaling pathways and/or enzymes involved in prostaglandin synthesis.     

Control of reproductive processes by growth hormone: extra- and intracellular mechanisms

Recent data on the association between growth hormone (GH) and male and female reproductive processes, as well as the effects of GH on these processes and on some reproductive and non-reproductive disorders, and possible extra- and intracellular mediators of its action are reviewed. The available data suggest that GH is an important endocrine and autocrine/paracrine regulator of reproduction. It controls proliferation, apoptosis, growth and differentiation and the secretory and generative activities of different reproductive organs. It also regulates their response to gonadotrophin-releasing hormone (GnRH) and gonadotropins. Despite the effects of GH on the IGF/IGFBP (insulin-like growth factor binding protein) system, oxytocin, steroids, activin, gonadotropin and gonadotropin receptors, the majority of GH's actions on the reproductive processes are probably mediated not by these substances but by specific GH receptors acting through cAMP/protein kinase A, protein kinase G, tyrosine kinase-, MAP kinase and CDC2 kinase-dependent intracellular mechanisms. Although GH treatments can increase the risk of some reproductive and non-reproductive disorders, they may be useful in improving gonadal function, inducing superovulation and in embryo production.     

Neuroregulation of growth hormone secretion in domestic animals

Growth hormone (GH) is essential for postnatal somatic growth, maintenance of lean tissue at maturity in domestic animals and milk production in cows. This review focuses on neuroregulation of GH secretion in domestic animals. Two hormones principally regulate the secretion of GH: growth hormone-releasing hormone (GHRH) stimulates, while somatostatin (SS) inhibits the secretion of GH. A long-standing hypothesis proposes that alternate secretion of GHRH and SS regulate episodic secretion of GH. However, measurement of GHRH and SS in hypophysial-portal blood of unanesthetized sheep and swine shows that episodic secretion of GHRH and SS do not account for all episodes of GH secreted. Furthermore, the activity of GHRH and SS neurons decreases after steers have eaten a meal offered for a 2-h period each day (meal-feeding) and this corresponds with reduced secretion of GH. Together, these data suggest that other factors also regulate the secretion of GH. Several neurotransmitters have been implicated in this regard. Thyrotropin-releasing hormone, serotonin and gamma-aminobutyric acid stimulate the secretion of GH at somatotropes. Growth hormone releasing peptide-6 overcomes feeding-induced refractoriness of somatotropes to GHRH and stimulates the secretion of GHRH. Norepinephrine reduces the activity of SS neurons and stimulates the secretion of GHRH via alpha(2)-adrenergic receptors. N-methyl-D,L-aspartate and leptin stimulate the secretion of GHRH, while neuropeptide Y stimulates the secretion of GHRH and SS. Activation of muscarinic receptors decreases the secretion of SS. Dopamine stimulates the secretion of SS via D1 receptors and inhibits the secretion of GH from somatotropes via D2 receptors. Thus, many neuroendocrine factors regulate the secretion of GH in livestock via altering secretion of GHRH and/or SS, communicating between GHRH and SS neurons, or acting independently at somatotropes to coordinate the secretion of GH.     

Effects of exogenous progesterone and/or prolactin on Haemonchus contortus infections in ovariectomized ewes

Reproduction can alter the course of ovine nematodiasis; fecal nematode egg concentrations often increase near lambing and throughout lactation, a phenomenon referred to as the periparturient rise. To identify the host mechanisms that might link these disparate events, i.e. lactation and the fecundity of gastrointestinal trichostrongyles, ovariectomized ewes were injected daily with progesterone and/or prolactin, or saline. Progesterone treatment commenced 20 days before inoculation with Haemonchus contortus and prolactin was administered throughout the 30-day infection period. Ovariectomized ewes receiving prolactin during the experimental infection maintained higher daily fecal egg concentrations than the progesterone, progesterone/prolactin, or the control treatment groups. However, ovariectomized ewes that received 20 days of pre-inoculation exposure to progesterone, as well as prolactin during the infection, had greater numbers of nematodes that were larger than the other treatment groups. Thus, the sequential delivery of these hormones that are associated with the reproductive cycle in ewes produced some of the same results that occur during periparturient rise.     

γ-氨基丁酸(GABA)的生殖生理作用

γ－氨基丁酸（GABA）作为一种抑制性神经递质，通过下丘脑－垂体－性腺轴本和性腺生理机能，从而参与激素的分泌调节。GABA对卵巢颗粒细胞孕酮的分泌调节随动情期不同而呈现抑制和促进作用。GABA也可能过多巴胺抑制系统抑制垂体激素LH和PRL的分泌。GABA与孕酮协同作用，能明显促进精子的获能，提高精子的体外能力。     

Reproductive Biology of Chamaeleo Pumilus Pumilus with Special Reference to the Role of the Corpus Luteum and Progesterone

The morphology of the female reproductive tract and of the ovaries of the ovoviviparous C. pumilus was studied. Sperm storage organs were found in the vaginal region and changes in the histological appearance of the corpus luteum indicated a cyclic function similar to that found in mammals. Studies with ¹⁴C labelled leucine suggested that amino acid transfer occurred between the mother and the embryos. A radio-immuno assay of the progesterone content of the corpus luteum showed that the corpus luteum contains 56,8 μg progesterone/g of fresh luteal tissue and that the mean plasma levels of progesterone increased from 945,6 pg/ml in non-gravid to 2296,0 pg/ml in gravid females. There also appears to be a post-ovulatory surge of progesterone (4946,0 pg/ml). Oestrogens were found to stimulate the oviducal mucosa while progesterone acted synergjstically with oestrogen. Oestrogens also stimulated the production of serum protein fractions. Progesterone caused yolk regression and thus prevented the onset of the next follicular phase. Studies involving ovariectomy and progesterone replacement, indicated that progesterone and the corpus luteum are essential for the maintenance of gestation. It was concluded that several features of the reproductive biology of C. pumilus can be considered to be distinctly mammalian and that these features are of considerable evolutionary importance.     

Basis of melengestrol acetate action as a progestin

To provide insight into potential mechanisms contributing to the various biological responses of cattle to treatment with progesterone, norgestomet, and melengestrol acetate (MGA), MCF-7 cells were utilized to determine the relative binding affinity of the progesterone receptor for MGA, norgestomet, progesterone, and a progesterone agonist (R5020), and to determine if progesterone, MGA, or norgestomet have estrogenic and/or anti-estrogenic activities. The progesterone receptor had greater affinity (P<0.05) for MGA, R5020, and norgestomet than for progesterone; and the affinity for norgestomet exceeded (P<0.05) that of MGA and R5020. Estrogen stimulates proliferation of MCF-7 cells; therefore these cells have been utilized as a bioassay to detect estrogenic and anti-estrogenic activity. Progesterone (10(-11) to 10(-5)M) did not promote cellular proliferation. However, MGA (10(-8), 10(-7), and 10(-6)M) increased (P<0.05) cell proliferation compared to the control group (10(-11) to 10(-9) and 10(-5)M MGA did not stimulate cell proliferation), and MGA-induced cell proliferation (10(-8)M) was reduced (P=0.095) by an estradiol antagonist (ICI 182,780; ICI). Cellular proliferation increased (P<0.05) with norgestomet (10(-5)M) compared to the control group (10(-11) to 10(-6)M norgestomet did not stimulate cell proliferation) and the increased proliferation was decreased (P<0.05) by ICI. Neither progesterone nor MGA demonstrated anti-estrogenic activity. Norgestomet (10(-10) to 10(-6)M) did reduce (P<0.05) maximal estrogen-stimulated cell proliferation, but not to basal levels. In summary, the affinities of the progesterone receptor for norgestomet, MGA, and progesterone are consistent with their effective dose to inhibit ovulation in vivo, but their progestin and their estrogenic/anti-estrogenic activities cannot fully explain why progesterone and norgestomet are more capable of reprogramming the reproductive axis in anestrous postpartum cows compared to MGA.     

The critical roles of progesterone receptor (PGR) in ovulation, oocyte developmental competence and oviductal transport in mammalian reproduction

Progesterone is critical for successful ovulation in the ovary and for the multi-faceted role of the oviduct in mammalian reproduction. Its effects are mediated by progesterone receptor (PGR), which is highly expressed in the ovary, specifically granulosa cells of preovulatory follicles in response to the luteinizing hormone (LH) surge that occurs just prior to ovulation, and in the oviduct, predominantly luminal epithelial cells but also muscle cells. This review will summarize research which shows that progesterone, via the actions of PGR, plays a key role in the functions of these cells and in the important periovulatory events of oocyte release, acquisition of oocyte developmental competence and oviductal transport of the newly formed embryo. PGR is a nuclear receptor that regulates the expression of many downstream target genes. However, although much is known about its expression characteristics in ovarian and oviductal cells, there is still much to unravel about the mechanisms by which PGR exerts its control over these important reproductive processes, particularly in the oviduct.     

Progesterone production by cultured luteal cells in the presence of bovine low- and high-density lipoproteins purified by heparin affinity chromatography.

The objectives of this study were to separate plasma lipoprotein particles based on the presence (low-density lipoproteins; LDL) or absence of apolipoprotein B (high-density lipoproteins; HDL) and to compare the abilities of bovine LDL and HDL to stimulate progesterone production by bovine luteal cells in culture. Plasma lipoproteins were isolated by ultracentrifugation and separated into LDL and HDL by heparin affinity chromatography. Luteal cultures were treated with LDL or HDL cholesterol for 48 h on d 3 of the culture (d 0 = day of dissociation). Progesterone production by luteal cells increased in a dose-dependent manner with increasing concentrations of either LDL or HDL cholesterol. There were no differences in the ability of LDL or HDL cholesterol to stimulate luteal cells to produce progesterone. Because LDL and HDL were equally potent, and experiment was designed to investigate the ability of modified LDL or reconstituted particles without apolipoproteins to stimulate progesterone production. Stimulation of luteal cell progesterone production by lysine-modified LDL was 70% of unmodified LDL. Progesterone production in the presence of phosphatidylcholine liposomes or BSA containing cholesterol was 50 and 108% of that obtained with HDL or LDL. Evidence indicated that apolipoprotein-free particles that contained free cholesterol but not cholesterol esters stimulated luteal cells to produce progesterone.     

Use of Equine Chorionic Gonadotropin to Control Reproduction of the Dairy Cow: A Review

Equine chorionic gonadotropin (eCG) is a member of the glycoprotein family of hormones along with LH, FSH and thyroid‐stimulating hormone. In non‐equid species, eCG shows high LH‐ and FSH‐like activities and has a high affinity for both FSH and LH receptors in the ovaries. On the granulosa and thecal cells of the follicle, eCG has long‐lasting LH‐ and FSH‐like effects that stimulate oestradiol and progesterone secretion. Thus, eCG administration in dairy cattle results in fewer atretic follicles, the recruitment of more small follicles showing an elevated growth rate, the sustained growth of medium and large follicles and improved development of the dominant and pre‐ovulatory follicle. In consequence, the quality of the ensuing CL is improved, and thereby progesterone secretion increased. Based on these characteristics, eCG treatment is utilized in veterinary medicine to control the reproductive activity of the cow by i) improving reproductive performance during early post‐partum stages; ii) increasing ovulation and pregnancy rates in non‐cyclic cows; iii) improving the conception rate in cows showing delayed ovulation; and finally, iv) eCG is currently included in protocols for fixed‐time artificial insemination since after inducing the synchrony of ovulation using a progesterone‐releasing device, eCG has beneficial effects on embryo development and survival. The above effects are not always observed in cyclic animals, but they are evident in animals in which LH secretion and ovarian activity are reduced or compromised, for instance, during the early post‐partum period, under seasonal heat stress, in anoestrus animals or in animals with a low body condition score.     

家畜视黄醇结合蛋白4（RBP4）的研究进展

视黄醇结合蛋白4（retinol-binding proteins 4,RBP4）是由孕酮受体产生的重要蛋白质,其促进转化生长因子（TGF）的表达,参与胚胎的早期发育。此外,RBP4还具有参与VA转运等功能。RBP4基因是影响家畜繁殖性能的候选基因,现已在绵羊、猪、牛和其他家畜中进行了广泛研究。综述了家畜RBP4基因的作用和近年来国内外的相关研究进展。     

Pregnancy recognition signaling mechanisms in ruminants and pigs

Maternal recognition of pregnancy refers to the requirement for the conceptus (embryo and its associated extra-embryonic membranes) to produce a hormone that acts on the uterus and/or corpus luteum (CL) to ensure maintenance of a functional CL for production of progesterone; the hormone required for pregnancy in most mammals. The pregnancy recognition signal in primates is chorionic gonadotrophin which acts directly on the CL via luteinizing hormone receptors to ensure maintenance of functional CL during pregnancy. In ruminants, interferon tau (IFNT) is the pregnancy recognition signal. IFNT is secreted during the peri-implantation period of pregnancy and acts on uterine epithelia to silence expression of estrogen receptor alpha and oxytocin receptor which abrogates the oxytocin-dependent release of luteolytic pulses of prostaglandin F2-alpha (PGF) by uterine epithelia; therefore, the CL continues to produce progesterone required for pregnancy. Pig conceptuses secrete interferon delta and interferon gamma during the peri-implantation period of pregnancy, but there is no evidence that they are involved in pregnancy recognition signaling. Rather, pig conceptuses secrete abundant amounts of estrogens between Days 11 to 15 of pregnancy required for maternal recognition of pregnancy. Estrogen, likely in concert with prolactin, prevents secretion of PGF into the uterine venous drainage (endocrine secretion), but maintains secretion of PGF into the uterine lumen (exocrine secretion) where it is metabolized to a form that is not luteolytic. Since PGF is sequestered within the uterine lumen and unavailable to induce luteolysis, functional CL are maintained for production of progesterone. In addition to effects of chorionic gonadotrophin, IFNT and estrogens to signal pregnancy recognition, these hormones act on uterine epithelia to enhance expression of genes critical for growth and development of the conceptus.     

Effect of reproductive status on intraocular pressure in cats

OBJECTIVE: To measure intraocular pressure (IOP) and progesterone concentrations in cats and to examine their reproductive organs to determine whether reproductive status affects IOP in cats. ANIMALS:75 sexually intact domestic shorthair cats scheduled to be neutered, including 28 males, 21 females not in estrus, 13 females in estrus, and 13 pregnant females. PROCEDURES: Applanation tonometry was conducted to measure IOP and radioimmunoassay was used to determine progesterone concentrations. Reproductive organs were examined at time of surgery. RESULTS: The IOP in female cats that were in estrus was significantly higher than IOP in female cats that were not in estrus. Progesterone concentrations significantly affected IOP in pregnant cats. CONCLUSIONS AND CLINICAL RELEVANCE: In cats, IOP is affected by changes in reproductive status. Such changes should be considered when interpreting tonometry results in this species.     

Corpus Luteum–Endometrium–Embryo Interactions in the Dairy Cow: Underlying Mechanisms and Clinical Relevance

Conception rates of dairy cows are currently declining at an estimated 1% every year. Approximately, 35% of embryos fail to prevent luteolysis during the first three weeks of gestation. Interactions between the corpus luteum, endometrium and embryo are critical to the successful establishment of pregnancy and inadequacies will result in the mortality of the embryo. For example, as little as a one day delay in the post-ovulatory rise of progesterone has serious consequences for embryo development and survival. Recently, we found that LH support, degree of vascularization and luteal cell steroidogenic capacity were not the major factors responsible for this luteal inadequacy, but are nevertheless essential for luteal development and function. Progesterone acting on its receptor in the endometrium stimulates the production of endometrial secretions on which the free-living embryo is dependent. However, their exact composition and effects of inadequate progesterone remains to be determined. The embryo is recognized through its secretion of interferon tau (IFNT), which suppresses luteolytic pulses of prostaglandin F_2α. In the cow, it is most likely that IFNT inhibits oxytocin receptor up-regulation directly and does not require the prior inhibition of oestrogen receptor α (ESR1). Unravelling the precise luteal-endometrium and embryo interactions is essential for us to understand pregnancy establishment and development of strategies to reverse the declining fertility of dairy cows.     

Production of progesterone and testosterone in ovarian granulosa cells in sows after administration of nonapeptide hormones in vitro]

In the present paper the effects of nonapeptide hormones and of some of their chemical analogues were investigated on progesterone and testosterone production in granulosa cells of sow ovaries; the experiments were made in vitro. This objective was given by data on potential regulatory roles of nonapeptides at the level of hypothalamus, pituitary and reproductive organs. The goal of this experiment was to analyze the effects of various doses of oxytocin (OT), arginine-8-vasopressin (AVP), arginine-8-vasotocin and of some of their analogues on progesterone and testosterone production in vitro in granulosa cells of sow ovaries. The production activity of granulosa cells was investigated which were obtained from slaughtered sows without any changes in their reproductive process and abnormalities in their reproductive organs. Follicles of the size 2-5 mm without marked paleness in the early follicular phase were selected for aspiration. Granulosa cells with determined viability (more than 75%) and concentration (2 million/ml) were cultivated in defined culture conditions (37.5 degrees C, 5% CO2) after threefold resuspension and centrifugation of follicle fluid. These hormonal preparations were used in the experiments: pFSH, synthetic OT, synthetic AVP, synthetic AVP with antidiuretic effects and synthetic AVT. Progesterone and testosterone concentrations were analyzed radioimmunoanalytically using commercial kits of the Institute of Radio ecology and Nuclear Technology at Kosice. Statistically significant differences between the groups were evaluated by Student's t-test. The administered preparations were found to influence progesterone and testosterone production in dependence on the doses applied (Figs. 1-6). OT stimulation of progesterone production in granulosa cells indicated its regulatory role in relation to secretion of this hormone.(ABSTRACT TRUNCATED AT 250 WORDS)