Hypothalamic gonadotropin-releasing hormone receptor activation stimulates oxytocin release from the rat hypothalamo-neurohypophysial system while melatonin inhibits this process

The present study was undertaken to investigate the influence of gonadotropin-releasing hormone (GnRH) and its agonist and antagonist on oxytocin (OT) release from the rat hypothalamo-neurohypophysial (H-N) system. An additional aim was to determine whether the possible response of oxytocinergic neurons to these peptides could be modified by melatonin through a cAMP-dependent mechanism.The results show that the highly selective GnRH agonist (i.e., [Des-Gly¹⁰,d-His(Bzl)⁶,Pro-NHEt⁹]-LHRH; Histrelin) stimulates the secretion of OT from an isolated rat H-N system. Melatonin significantly inhibited basal and histrelin-induced release of OT in vitro, and displayed no significant influence on OT release in the presence of GnRH or its antagonist. Addition of melatonin to a medium containing forskolin resulted in significant reduction of OT secretion from the H-N system. On the other hand, addition of forskolin to a medium containing both histrelin and melatonin did not further alter the inhibitory influence of melatonin on the histrelin-dependent secretion of OT in vitro. Intracerebroventricular (icv) infusion (experiment in vivo) of a GnRH antagonist resulted in substantial inhibition of OT release, thus revealing the stimulatory action of endogenous GnRH. In melatonin-treated animals, blood plasma OT levels were not changed in comparison to the vehicle.Our present data strongly suggests that activation of the GnRH receptor in the hypothalamus is involved in stimulation of OT secretion from the rat H-N system. It has also been shown, under experimental in vitro conditions, that melatonin fully suppresses the response of oxytocinergic neurons to the GnRH agonist - histrelin. The effect of melatonin on OT release is mediated by the cAMP-dependent mechanism, although other mechanisms of action are also possible.     

Gonadotropin response to synthetic gonadotropin hormone-releasing hormone (GnRH) in chronic schizophrenia

GnRH stimulation tests were performed in 15 adult male chronic hebephrenic schizophrenics and 15 oligophrenic controls, matched for age and length of hospitalization. GnRH was given at doses of 50, 100 and 150 gamma to five subjects of each type, and FSH and LH levels in the blood were assayed at 0, 10, 20, 30, 60, and 90 minutes. The tests were performed twice in schizophrenics off therapy and after 10, 20 and 30 days of chlorpromazine therapy (4 mg/kg body weight/day, peros). The controls were not given chlorpromazine and were tested only twice. Schizophrenics showed relative increases in both FSH and LH which were greater than those of the controls, and the response persisted longer. Chlorpromazine had no effect on the test.     

Corticotropin-releasing factor (CRF) inhibits gonadotropin-releasing hormone (GnRH) release from superfused rat hypothalami in vitro

A dose-dependent suppression of basal gonadotropin-releasing hormone (GnRH) release was observed when rat hypothalamic slices were superfused with human/rat corticotropin-releasing factor (CRF) (10(-12) to 10(-8) M). CRF was also found to significantly reduce the amount of GnRH released in response to 56 mM KCl. These in vitro results demonstrate that the CRF inhibition of GnRH secretion observed in vivo occurs within the hypothalamus and independently of the CRF activation of the pituitary-adrenal-gonadal axis.     

Gonadotrophin-releasing hormone (GnRH) release in marmosets II: pulsatile release of GnRH and pituitary gonadotrophin in adult females

Unlike other mammals, including rodents, Old World primates and humans, common marmosets and probably all other New World primates synthesise and release chorionic gonadotrophin (CG), and not luteinising hormone (LH) from pituitary gonadotrophs. However, little is known about the physiological dynamics of gonadotrophin-releasing hormone (GnRH)-regulated CG release from gonadotrophs and whether such CG release has pulsatile release characteristics similar to those of LH in other mammalian species. Consequently, we performed a series of in vivo and in vitro studies in ovariectomised laboratory rats and female marmosets to compare GnRH-induced pituitary LH and CG release characteristics, respectively. Exogenous GnRH stimulated a slower onset of release of marmoset pituitary CG, both in vivo and in vitro, and induced an approximately 400% greater increase in the duration of marmoset pituitary CG release compared to that for rat LH. Not surprisingly, hypothalamic pulsatile release of GnRH in vivo was not obviously concordant with endogenous episodic changes in circulating levels of CG in marmosets, in contrast to the clear concordance observed between in vivo GnRH and LH release previously demonstrated in rats and other mammals. Pituitary CG release in marmosets thus demonstrates considerable divergence from the timely hypothalamic GnRH-regulated LH release in other female mammals, implying potentially different physiological dynamics in gonadotrophin regulation of marmoset ovarian function.     

Gonadotrophin-releasing hormone (GnRH) release in marmosets I: in vivo measurement in ovary-intact and ovariectomised females

In vivo hypothalamic gonadotrophin-releasing hormone (GnRH) release was characterised for the first time in a New World primate. A nonterminal and repeatable push-pull perfusion (PPP) technique reliably measured GnRH in conscious common marmoset monkeys. Nineteen adult females (n = 8 ovary-intact in the mid-follicular phase; n = 11 ovariectomised) were fitted with long-term cranial pedestals, and a push-pull cannula was temporarily placed in unique locations within the pituitary stalk-median eminence (S-ME) 2 days prior to each PPP session. Marmosets underwent 1-3 PPPs (32 PPPs in total) lasting up to 12 h. Plasma cortisol levels were not elevated in these habituated marmosets during PPP, and PPP did not disrupt ovulatory cyclicity or subsequent fertility in ovary-intact females. GnRH displayed an organised pattern of release, with pulses occurring every 50.0 +/- 2.6 min and lasting 25.4 +/- 1.3 min. GnRH pulse frequency was consistent within individual marmosets across multiple PPPs. GnRH mean concentration, baseline concentration and pulse amplitude varied predictably with anatomical location of the cannula tip within the S-ME. GnRH release increased characteristically in response to a norepinephrine infusion and decreased abruptly during the evening transition to lights off. Ovary-intact (mid-follicular phase) and ovariectomised marmosets did not differ significantly on any parameter of GnRH release. Overall, these results indicate that PPP can be used to reliably assess in vivo GnRH release in marmosets and will be a useful tool for future studies of reproductive neuroendocrinology in this small primate.     

Gonadotropin-releasing hormone immunoreactivity in the adult and fetal human olfactory system

Studies in fetal brain tissue of rodents, nonhuman primates and birds have demonstrated that cells containing gonadotropin-releasing hormone (GnRH) migrate from the olfactory placode across the nasal septum into the forebrain. The purpose of this study was to examine GnRH neurons in components of the adult and fetal human olfactory system. In the adult human brain (n=4), immunoreactive GnRH was evident within diffusely scattered cell bodies and processes in the olfactory bulb, olfactory nerve, olfactory cortex, and nervus terminalis located on the anterior surface of the gyrus rectus. GnRH-immunoreactive structures showed a similar distribution in 20-week human fetal brains (n=2), indicating that the migration of GnRH neurons is complete at this time. In 10-11-week fetal brains (n=2), more cells were noted in the nasal cavity than in the brain. Our data are consistent with observations made in other species, confirming olfactory derivation and migration of GnRH neurons into the brain from the olfactory placode.     

Gonadotropin-releasing hormone (GnRF), Molluscan cardioexcitatory peptide (FMRFamide), enkephalin and related neuropeptides affect goldfish retinal ganglion cell activity

Recently gonadotropin-releasing hormone (GnRF)-like and molluscan cardioexcitatory peptide (FMRFamide)-like compounds have been colocalized immunocytochemically to the terminal nerve, a presumed olfactoretinal efferent system in goldfish. In the present study these and related neuropeptides were shown to affect ganglion cell activity, recorded extracellularly, when applied to the isolated superfused goldfish retina. GnRF was usually excitatory. Salmon GnRF (sGnRF) was 10-30x more potent than chicken or mammalian GnRF. FMRFamide and enkephalin also were often excitatory but caused more varied responses than sGnRF. Met5-enkephalin-Arg6-Phe7-NH2 (YGGFMRFamide), which contains both enkephalin and FMRFamide sequences, tended to act like both of these peptides but with mainly enkephalin-like properties. Neuropeptide Y and the C-terminal hexapeptide of pancreatic polypeptides, whose C-terminus (-Arg-Tyr-NH2) is closely related to that of FMRFamide (-Arg-Phe-NH2), gave no consistent responses. Threshold doses were equivalent to: 0.1 microM for sGnRF; 0.5 microM for YGGFMRFamide; 1.5 microM for FMRFamide and enkephalin. Rapid, complete and irreversible desensitization was induced by single, 10-20x threshold doses of sGnRF; but desensitization was infrequent and limited with the other peptides. In general, all peptides tested affected the spatially and chromatically antagonistic receptive field components similarly, but selective actions were seen in a few cases with FMRFamide and with the opioid antagonist, naloxone. Responses, especially to sGnRF and FMRFamide, tended to be most frequently obtained and pronounced in winter and spring, suggesting a correlation with seasonally regulated sexual and reproductive activity. Our observations provide further evidence for transmitter-like roles of neuropeptides related to sGnRF and FMRFamide in the teleostean terminal nerve. The actions of agonists and antagonists, singly and in combination, imply strongly that there are distinctive postsynaptic receptors and/or neural pathways for GnRF-, FMRFamide- and enkephalin-like peptides in the goldfish retina.     

Assessment of the effects of a synthetic gonadotropin-releasing hormone associated peptide on hormone release from the in situ and ectopic pituitaries in adult male rats

The in vivo effects of synthetic human gonadotropin-releasing hormone associated peptide (GAP) were evaluated in adult male rats. In normal rats, intracerebroventricular (III ventricle) injection of 5 ng GAP significantly increased plasma LH levels after 60 minutes. Intracerebroventricular administration of 5, 25 or 125 ng of GAP elevated circulating LH levels also at 120 minutes of injection but did not alter plasma FSH, prolactin or testosterone concentrations. In hypophysectomized-pituitary-grafted rats injection of 125 ng GAP directly into the ectopic pituitary induced no changes in peripheral hormone levels. However, intrapituitary graft injection of 25 ng of GnRH significantly elevated circulating levels of LH and testosterone. These results indicate that the ectopic pituitary graft can respond to acute exogenous GnRH stimulation and that the commercially available synthetic GAP fails to inhibit prolactin release in adult male rats.     

Gonadotropin-releasing hormone within the organum vasculosum lamina terminalis in the ovariectomized, estrogen/progesterone-treated rat: A quantitative immunocytochemical study using image analysis

No specific function has been ascribed to the high gonadotropin-releasing hormone (GnRH) content of the organum vasculosum lamina terminalis (OVLT). The objective of this study was to determine whether levels of GnRH within the OVLT are altered during cyclic gonadotropin secretion. GnRH levels were determined at various times during an estrogen/progesterone (E/P)-induced LH surge. Groups of E/P and sesame oil-treated animals were decapitated at 12.00 h, 14.00 h, 16.00 h, 18.00 h, and 22.00 h following the P or oil treatment. Morphological localization, as well as quantitation of immunoreactive GnRH within discrete regions of the brain was achieved by combining unlabeled antibody immunocytochemistry with computerized image analysis. Analysis of GnRH levels in the OVLT revealed that at any of the 5 times examined, there was: (1) no significant difference among controls, (2) no significant difference among E/P-treated animals, and (3) no significant difference between E/P-treated versus control animals. In contrast, ME GnRH levels in E/P-treated rats showed the expected decrease prior to the onset of the LH surge. These findings suggest that the level of GnRH detected in axon terminals within the OVLT cannot be related directly to the serum LH status of ovariectomized, E/P-treated rats. It is therefore possible that GnRH within the OVLT might function in a neuromodulatory role, rather than as a direct regulator of cyclic gonadotropin secretion.     

Gonadotropin-releasing hormone neurons, NMDA receptors, and their regulation by steroid hormones across the reproductive life cycle

The effects of ovarian steroid hormones on gonadotropin-releasing hormone (GnRH) neurons have been studied for many years. In addition to their regulation by sex steroids, GnRH neurons are affected by inputs from neurotransmitters such as glutamate, acting via the NMDA receptor (NMDAR). Moreover, the NMDAR itself is subject to estrogen regulation. Thus, effects of ovarian steroids on GnRH neurons and the NMDAR, and their interactions, are under intense investigation. Messenger RNA and protein levels of GnRH and NMDAR subunits were measured in neuroendocrine brain regions in response to estrogen treatment, or across the reproductive cycle. Stimulatory effects of ovarian steroids on GnRH gene expression occur during the preovulatory LH surge in young adult rats, and this is abolished in middle-aged rats that have an attenuated LH surge. Effects of estrogen on GnRH neurons have also been studied in the ovariectomized, estrogen-primed rat, and while results vary between laboratories, there appear to be age-related changes in the sensitivity of GnRH neurons to estrogen. Estrogen also has effects on NMDAR mRNA levels. In intact rats, mRNA levels of NMDAR decrease during reproductive aging in the preoptic area, the site of GnRH perikarya, while in the medial basal hypothalamus-median eminence, the site of GnRH neuroterminals, levels of NMDAR subunit mRNAs increase with aging. Thus, glutamatergic inputs to GnRH perikarya and neuroterminals and other neuroendocrine cells may change during reproductive aging in intact rats. In ovariectomized rats, NMDAR subunit mRNA levels also undergo age-related changes, and respond to estrogen replacement in a subunit- and age-specific manner. Notably, there are major differences in NMDAR gene expression during aging between intact and ovariectomized rats, suggesting that ovarian factors other than estrogen play a role in the regulation of this receptor.     

Polycystic ovary syndrome: Understanding the role of the brain

Polycystic ovary syndrome (PCOS) is a prevalent endocrine disorder and the leading cause of anovulatory infertility. Characterised by hyperandrogenism, menstrual dysfunction and polycystic ovaries, PCOS is a broad-spectrum disorder unlikely to stem from a single common origin. Although commonly considered an ovarian disease, the brain is now a prime suspect in both the ontogeny and pathology of PCOS. We discuss here the neuroendocrine impairments present in PCOS that implicate involvement of the brain and review evidence gained from pre-clinical models of the syndrome about the specific brain circuitry involved. In particular, we focus on the impact that developmental androgen excess and adult hyperandrogenemia have in programming and regulating brain circuits important in the central regulation of fertility. The studies discussed here provide compelling support for the importance of the brain in PCOS ontogeny and pathophysiology and highlight the need for a better understanding of the underlying mechanisms involved.     

脑膜瘤中EGFR和PTEN的表达及意义

目的探讨脑膜瘤组织中表皮生长因子受体（EGFR）和人10号染色体上缺失的磷酸酶和张力蛋白类似物（PTEN）表达的相互关系及意义。方法应用免疫组织化学SP法对60例脑膜瘤标本的EGFR和PTEN表达进行检测与分析。结果在90％（54／60）的脑膜瘤中EGFR过表达，非典型脑膜瘤及间变性脑膜瘤中EGFR的染色强度明显高于典型脑膜瘤（P〈0．05）；PTEN在各病理级别脑膜瘤中的阳性率为100％，而且各病理级别脑膜瘤中PTEN的染色强度无统计学差别（P〉0．05）。结论EGFR的表达强度与脑膜瘤的恶性程度有关，抑癌基因PTEN在脑膜瘤中未见缺失，可能与脑膜瘤的发生、发展无关。     

Developmental changes in the mammalian gonadotropin-inhibitory hormone (GnIH) ortholog RFamide-related peptide (RFRP) and its cognate receptor GPR147 in the rat hypothalamus

The mammalian gonadotropin-inhibitory hormone (GnIH) ortholog RFamide-related peptide (RFRP) is considered to act on gonadotropin-releasing hormone (GnRH) neurons and on the pituitary to inhibit gonadotropin release and synthesis. To understand the functional significance of this neuropeptide, we investigated the physiological changes in RFRP at mRNA and peptide levels, as well as at the mRNA level of its cognate receptor, G protein-coupled receptor 147 (GPR147), in the rat hypothalamus during development. We also investigated the effects of gonadal steroids on mRNA expression levels of these molecules. In male rats, mRNA expressions of both RFRP and GPR147 increased from postnatal days 12 and 16, peaking at postnatal days 35 and 42, respectively. However, their expressions fell at postnatal day 49. In female rats, mRNA expression of RFRP continued to increase throughout development; mRNA expression of GPR147 in female rats increased from postnatal day 16, peaking at postnatal day 28, but decreased from postnatal day 35. The hypothalamic contents of RFRP on postnatal days 28 and 42 were significantly higher than on postnatal day 4 in male rats, and those on postnatal day 42 were significantly higher than those on postnatal days 4 and 28 in females. Neither orchidectomy nor ovariectomy influenced mRNA expression levels of RFRP or GPR147 in the prepubertal period when endogenous sex steroid levels were low in males and females. Administration of estradiol-17β (E2) increased mRNA expression of RFRP in prepubertal females. These results suggest that the hypothalamic RFRP system changes during development. An ovarian sex steroid, E2, may stimulate mRNA expression of RFRP in the prepubertal period when the basal E2 concentration is low.     

Gonadotropin-releasing hormone (GnRH) neuron migration: Initiation, maintenance and cessation as critical steps to ensure normal reproductive function

GnRH neurons follow a carefully orchestrated journey from their birth in the olfactory placode area. Initially, they migrate along with the vomeronasal nerve into the brain at the cribriform plate, then progress caudally to sites within the hypothalamus where they halt and send projections to the median eminence to activate pituitary gonadotropes. Many factors controlling this precise journey have been elucidated by the silencing or over-expression of candidate genes in mouse models. Importantly, a number of these factors may not only play a role in normal physiology of the hypothalamic-pituitary-gonadal axis but also be mis-expressed to cause human disorders of GnRH deficiency, presenting as a failure to undergo normal pubertal development. This review outlines the current cadre of candidates thought to modulate GnRH neuronal migration. The further elucidation and characterization of these factors that impact GnRH neuron development may shed new light on human reproductive disorders and provide potential targets to develop new pro-fertility or contraceptive agents.     

Induction of Galanin mRNA in GnRH Neurons by Estradiol and its Facilitation by Progesterone

On the day of proestrus in the rat, rising plasma levels of estradiol (E) act in concert with progesterone (P) to trigger a preovulatory release of gonadotropins. Cellular levels of galanin mRNA in GnRH neurons are increased in association with the proestrous surge of gonadotropin secretion; however, the relative contribution made by E and P to the induction of galanin mRNA expression in GnRH neurons is unknown. We investigated the role of E and P in the induction of galanin gene expression in GnRH neurons by examining the effects of different combinations of E (estradiol benzoate; 50 μg and P (5 mg)) on the LH surge and the concomitant induction of galanin mRNA in GnRH neurons. We sacrificed ovariectomized adult rats after 1 of 4 treatments: Group 1: vehicle control (n=6); Group 2: P alone (n=7) Group 3: E alone (n=7); Group 4: combined E/P (n=6); the animals were killed at 18.00 h at the time of the LH surge. The brains from these animals were processed by double-label in situ hybridization to allow measurement of galanin mRNA levels in GnRH neurons. GnRH neurons were identified with a digoxigenin-labeled cRNA probe for GnRH mRNA, and galanin mRNA was detected and measured simultaneously with an ³⁵S-labeled cRNA probe coupled with computerized grain counting. Estimation of cellular levels of GnRH mRNA was accomplished with single-label in situ hybridization, an ³⁵S-labeled GnRH cRNA probe and computerized grain counting. We observed a 3-fold induction of galanin mRNA in the GnRH neurons of animals treated with E alone compared with those treated with the vehicle alone (vehicle: 13±2 vs E: 42±4 grains/cell (g/c); P<0.01); LH levels in the E-treated animals were elevated, albeit moderately, with respect to the vehicle controls. Compared with vehicle-treated animals, those treated with the combination of E and P showed a 5-fold induction of galanin mRNA in GnRH neurons (68±9 g/c), which was significantly (P<0.01) greater than that observed in the animals treated with E alone; in addition, the magnitude of the LH surge was much greater (P<0.05) in the E/P-treated group compared with the E alone group. In contrast, compared to the vehicle controls, animals treated with P alone (15±2 g/c) showed no discernable effect on galanin mRNA levels; moreover, no LH surge occurred in the P alone group. Neither the number of identified GnRH cells nor their content of GnRH mRNA differed significantly among the experimental groups (GnRH mRNA signal: vehicle controls: 153±6 vs E: 159±6 vs E/P: 153±3 vs P: 148±8 g/c). We conclude that while E is the primary ovarian signal inducing galanin mRNA expression in GnRH neurons and the LH surge itself, P plays a facilitatory role in both of these processes.     

Gonadotropin-releasing hormone (GnRH) immunoreactive system in the brain of the dwarf gourami (Colisa lalia) as revealed by light microscopic immunocytochemistry using a monoclonal antibody to common amino acid sequence of GnRH

The present paper aims to give a morphological basis for the study of the terminal nerve system and its relation to the whole gonadotropin-releasing hormone (GnRH) immunoreactive (ir) neuronal system. We examined the GnRH-ir neuronal system of a tropical fish, the dwarf gourami, by using a recently developed monoclonal antibody against GnRH (LRH13) which recognizes the amino acid sequence common to all known variants of GnRH (Park and Wakabayashi, Endocrinol. Jpn. 33:257-272, '86). The ganglion cells of the terminal nerve (TN-ggl cells) in the transitional area between the olfactory bulb and the telencephalon reacted strongly with the LRH13. A distinct bundle of axons emanating from the TN-ggl cells ran caudally through the ventral telencephalon and the preoptic area. Some of these axons entered the optic nerve and innervated the retina. The remaining axons continued caudally to enter the hypothalamus and the midbrain. A second group of GnRH-ir cell bodies was found in the preoptic area. A distinct bundle of GnRH-ir fibers originating from these cell bodies innervated the pituitary. This pathway is equivalent to the preoptico-infundibular pathway of other vertebrates, and the GnRH in this pathway is presumed to function as hypophysiotrophic hormone to facilitate the release of gonadotropins from the pituitary. The distribution of GnRH-ir fibers in the brain was extensive. Most fibers apparently originated from the TN-ggl cells and covered various brain regions from the olfactory bulb to the spinal cord. They were especially abundant in the olfactory bulb, ventral telencephalon, preoptic area, optic tectum, and some hypothalamic areas. Thus, GnRH might function as a neuromodulator and/or neurotransmitter in these areas. The abundant GnRH-ir fibers in the ventral telencephalon and the preoptic area might affect some aspects of sexual behavior, since these areas have been suggested to be involved in the control of sexual behavior in teleosts.