Hypothalamic Gonadotropin-releasing Hormone Pulse Generator Research Articles

Investigation of irisin's role in pubertal onset physiology in female rats

ObjectiveThe timing of pubertal development is closely related to metabolic status and energy reserves. It is thought that irisin, which is involved in the regulation of energy metabolism and is shown to be present in the hypothalamo-pituitary-gonadal (HPG) axis, may play a role in this process. In our study, we aimed to investigate the effect of irisin administration on pubertal development and HPG axis in rats. Design-methods36 female rats were included in the study were divided into 3 groups: 100 ng/kg/day irisin treatment group (irisin-100), 50 ng/kg/day irisin treatment group (irisin-50), and control group. On the 38th day, serum samples were taken to determine levels of luteinizing hormone (LH) and follicle stimulating hormone (FSH), estradiol and irisin. Brain hypothalamus samples were taken to determine levels of pulsatile gonadotropin-releasing hormone (GnRH), kisspeptin, neurokinin-B, dynorphin (Dyn), and makorin ring finger protein-3 (MKRN3). ResultsVaginal opening and estrus were seen firstly in the irisin-100 group. At the end of the study, the highest rate of vaginal patency was found in the irisin-100 group. Hypothalamic protein expression levels of GnRH, NKB and Kiss1 in homogenates; serum FSH, LH, and estradiol levels were the highest in the irisin-100 group, followed by the irisin-50 and control groups, respectively. Ovarian sizes were significantly greater in the irisin-100 group compared to the other groups. The hypothalamic protein expression levels of MKRN3 and Dyn were the lowest in the irisin-100 group. ConclusionsIn this experimental study, irisin triggered the onset of puberty in a dose-dependent manner. Irisin administration caused the excitatory system to dominate in the hypothalamic GnRH pulse generator.

OR17-2 GABAergic Signalling in the Posterodorsal Medial Amygdala Mediates Psychological Stress-induced Suppression of the GnRH Pulse Generator in Female Mice

Abstract Psychological stress is linked to infertility and reproductive dysfunction by suppressing the hypothalamic gonadotropin-releasing hormone (GnRH) pulse generator and pulsatile luteinising hormone (LH) secretion. The posterodorsal subnucleus of the medial amygdala (MePD) is a key upstream regulator of GnRH pulse generator activity and displays increased neuronal activation in response to psychological stressors. The MePD is primarily a GABAergic nucleus with a strong GABAergic projection to hypothalamic reproductive centres, however, their functional significance has not been determined. We hypothesis that MePD GABAergic signalling is a crucial mediator of psychological stress-induced suppression of pulsatile LH secretion. We used a chemogenetic approach to selectively silence MePD GABA neurones during psychological stress to determine the effect on pulsatile LH secretion. Female ovariectomised Vgat-cre mice (n=7) were virally infected to selectively express inhibitory hM4DGi-DREADDs in MePD GABA neurones. Mice were exposed to either predator odour (PO) or restraint stress and given an intraperitoneal injection of either saline or the DREADD chemical activator clozapine-N-oxide (CNO, 5mg/kg). Statistical significance was determined using a one-way ANOVA and post-hoc Tukey test. Data are presented as mean ± SEM and p<0.05 was considered significant. PO exposure significantly increased LH inter-pulse interval (IPI) in the saline group (prestress: 19.05 ± 1.16 min vs. PO: 29.28 ± 3.16 min, p<0.05, n=7) and this effect was blocked by DREADD-mediated inhibition of MePD GABA neurones (prestress: 20.59 ± 1.12 min vs. PO: 21.31 ± 1.76 min, p>0.05, n=7). Restraint stress dramatically increased LH IPI in the saline group (prestress: 17.78 ± 1.02 min vs. restraint: 45.71 ± vs 5.50 min, p<0.05, n=7) and DREADD-mediated inhibition of MePD GABA neurones blocked this effect (prestress: 18.57 ± 1.97 min vs. restraint: 27.26 ± 4.02 min, p>0.05, n=7). Direct projections from the MePD to the GnRH pulse generator in the hypothalamic arcuate nucleus (ARC) have been identified, however, their phenotype is unknown. In this study, we optogenetically stimulated potential MePD GABAergic projection neurone terminals in the ARC and determined the effect on LH IPI. MePD GABA neurones in female Vgat-cre ovariectomised mice (n=6) were virally infected to express channelrhodopsin 2 protein and MePD GABAergic terminals in the ARC were selectively stimulated by blue light via a chronically implanted fibre-optic cannula in the ARC. Sustained optogenetic stimulation at 10 and 20Hz of MePD GABAergic terminals in the ARC dose-dependently suppressed pulsatile LH secretion (LH IPI; control: 23.34 ± 2.11 min vs. 10Hz: 38.08 ± 3.61 min; 20Hz: 56.67 ± 3.33 min, p<0.05, n=6). These findings confirm a functionally significant MePD GABAergic projection to the ARC and highlight the importance of GABA signalling in the MePD in mediating stress-induced suppression of reproductive function. Collectively, these findings provide novel insight into the amygdala neural circuitry underpinning stress-induced reproductive function. Presentation: Sunday, June 12, 2022 11:15 a.m. - 11:30 a.m.

Role of Posterodorsal Medial Amygdala Urocortin-3 in Pubertal Timing in Female Mice.

Post-traumatic stress disorder impedes pubertal development and disrupts pulsatile LH secretion in humans and rodents. The posterodorsal sub-nucleus of the medial amygdala (MePD) is an upstream modulator of the hypothalamic gonadotropin-releasing hormone (GnRH) pulse generator, pubertal timing, as well as emotional processing and anxiety. Psychosocial stress exposure alters neuronal activity within the MePD increasing the expression of Urocortin3 (Ucn3) and its receptor corticotropin-releasing factor type-2 receptor (CRFR2) while enhancing the inhibitory output from the MePD to key hypothalamic reproductive centres. We test the hypothesis that psychosocial stress, processed by the MePD, is relayed to the hypothalamic GnRH pulse generator to delay puberty in female mice. We exposed C57Bl6/J female mice to the predator odor, 2,4,5-Trimethylthiazole (TMT), during pubertal transition and examined the effect on pubertal timing, pre-pubertal LH pulses and anxiety-like behaviour. Subsequently, we virally infected Ucn3-cre-tdTomato female mice with stimulatory DREADDs targeting MePD Ucn3 neurons and determined the effect on pubertal timing and pre-pubertal LH pulse frequency. Exposure to TMT during pubertal development delayed puberty, suppressed pre-pubertal LH pulsatility and enhanced anxiety-like behaviour, while activation of MePD Ucn3 neurons reduced LH pulse frequency and delayed puberty. Early psychosocial stress exposure decreases GnRH pulse generator frequency delaying puberty while inducing anxiety-behaviour in female mice, an effect potentially involving Ucn3 neurons in the MePD.

Novel insights into the metabolic action of Kiss1 neurons.

Kiss1 neurons are essential regulators of the hypothalamic–pituitary–gonadal (HPG) axis by regulating gonadotropin-releasing hormone (GnRH) release. Compelling evidence suggests that Kiss1 neurons of the arcuate nucleus (Kiss1ARC), recently identified as the hypothalamic GnRH pulse generator driving fertility, also participate in the regulation of metabolism through kisspeptinergic and glutamatergic interactions with, at least, proopiomelanocortin (POMC) and agouti-related peptide (AgRP)/neuropeptide Y (NPY) neurons, located in close apposition with Kiss1ARC. This review offers a comprehensive overview of the recent developments, mainly derived from animal models, on the role of Kiss1 neurons in the regulation of energy balance, including food intake, energy expenditure and the influence of circadian rhythms on this role. Furthermore, the possible neuroendocrine pathways underlying this effect, and the existing controversies related to the anorexigenic action of kisspeptin in the different experimental models, are also discussed.

The neurobiological mechanism underlying hypothalamic GnRH pulse generation: the role of kisspeptin neurons in the arcuate nucleus.

This review recounts the origins and development of the concept of the hypothalamic gonadotropin-releasing hormone (GnRH) pulse generator. It starts in the late 1960s when striking rhythmic episodes of luteinizing hormone secretion, as reflected by circulating concentrations of this gonadotropin, were first observed in monkeys and ends in the present day. It is currently an exciting time witnessing the application, primarily to the mouse, of contemporary neurobiological approaches to delineate the mechanisms whereby Kiss1/NKB/Dyn (KNDy) neurons in the arcuate nucleus of the hypothalamus generate and time the pulsatile output of kisspeptin from their terminals in the median eminence that in turn dictates intermittent GnRH release and entry of this decapeptide into the primary plexus of the hypophysial portal circulation. The review concludes with an examination of questions that remain to be addressed.

The neurobiological mechanism underlying hypothalamic GnRH pulse generation: the role of kisspeptin neurons in the arcuate nucleus

This review recounts the origins and development of the concept of the hypothalamic gonadotropin-releasing hormone (GnRH) pulse generator. It starts in the late 1960s when striking rhythmic episodes of luteinizing hormone secretion, as reflected by circulating concentrations of this gonadotropin, were first observed in monkeys and ends in the present day. It is currently an exciting time witnessing the application, primarily to the mouse, of contemporary neurobiological approaches to delineate the mechanisms whereby Kiss1/NKB/Dyn (KNDy) neurons in the arcuate nucleus of the hypothalamus generate and time the pulsatile output of kisspeptin from their terminals in the median eminence that in turn dictates intermittent GnRH release and entry of this decapeptide into the primary plexus of the hypophysial portal circulation. The review concludes with an examination of questions that remain to be addressed.

Effects of puerarin on estrogen-regulated gene expression in gonadotropin-releasing hormone pulse generator of ovariectomized rats

Effects of puerarin on the hypothalamic gonadotropin-releasing hormone (GnRH) pulse generator function is investigated, for the first time, in ovariectomized rats at the level of mRNA expression of estrogen-responsive genes, e.g., estrogen receptor (ER), GnRH and its receptor (GnRHR). Rats were treated orally for 90 days either with a soy-free diet containing two different doses of puerarin (low dose of 600 mg/kg and high dose of 3000 mg/kg) or estradiol benzoate (E2B) at either low dose (4.3 mg/kg) or high dose (17.3 mg/kg). Levels of mRNA expression in the medial preoptic area/anterior hypothalamus (MPOA/AH), mediobasal hypothalamus/median eminence (MBH/ME) and adenohypophysis were measured by quantitative TaqMan® real-time RT-PCR. Plasma levels of luteinizing hormone (LH) and prolactin (PRL) were measured by radioimmunoassay. In the MPOA/AH, both puerarin and E2B decreased ERα mRNA levels without any significant changes in ERβ and GnRH mRNA levels. Both puerarin and E2B did not significantly alter the expression levels of ERα, ERβ and GnRHR in the MBH/ME. E2B exerted significant effects on the down-regulation of adenohypophyseal GnRHR mRNA transcripts and serum LH levels. Puerarin did not cause significant changes in pituitary GnRHR mRNA transcripts and serum LH and PRL levels. This is the first study to demonstrate that in ovariectomized rat models of ovarian hormone deprivation, puerarin acted as a weak estrogen-active compound in the hypothalamic GnRH pulse generator through the downregulation of MPOA/AH ERα mRNA expression.

Definition of the hypothalamic GnRH pulse generator in mice.

The pulsatile release of luteinizing hormone (LH) is critical for mammalian fertility. However, despite several decades of investigation, the identity of the neuronal network generating pulsatile reproductive hormone secretion remains unproven. We use here a variety of optogenetic approaches in freely behaving mice to evaluate the role of the arcuate nucleus kisspeptin (ARNKISS) neurons in LH pulse generation. Using GCaMP6 fiber photometry, we find that the ARNKISS neuron population exhibits brief (∼1 min) synchronized episodes of calcium activity occurring as frequently as every 9 min in gonadectomized mice. These ARNKISS population events were found to be near-perfectly correlated with pulsatile LH secretion. The selective optogenetic activation of ARNKISS neurons for 1 min generated pulses of LH in freely behaving mice, whereas inhibition with archaerhodopsin for 30 min suppressed LH pulsatility. Experiments aimed at resetting the activity of the ARNKISS neuron population with halorhodopsin were found to reset ongoing LH pulsatility. These observations indicate the ARNKISS neurons as the long-elusive hypothalamic pulse generator driving fertility.

Aging ovary and the role for advanced glycation end products

The hypothalamic gonadotropin-releasing hormone pulse generator, the pituitary gonadotropes, the ovaries, and the uterus play a crucial role in female fertility. A decline in reproductive performance represents a complex interplay of actions at all levels of the hypothalamic-pituitary-ovarian axis. Recently, in the field of female reproductive aging attention is drawn to the carbonyl stress theory. Advanced glycation end products (AGEs) contribute directly to protein damage, induce a chain of oxidative stress (OS) reactions, and increase inflammatory reactions. Here, we highlight some of the mechanisms underlying glycation damage in the ovary. Searches of electronic databases were performed. Articles relevant to possible role of OS, AGEs, and receptor for AGE (RAGE) in aging ovary were summarized in this interpretive literature review. Follicular microenvironment undergoes an increase in OS with aging. Data support the role of OS in ovulatory dysfunction because AGEs are well-recognized mediators of increased OS. RAGE and AGE-modified proteins with activated nuclear factor-kappa B are expressed in human ovarian tissue. It was suggested that accumulation of AGEs products at the level of the ovarian follicle might trigger early ovarian aging or could be responsible for reduced glucose uptake by granulosa cells, potentially altering follicular growth. Moreover, impaired methylglyoxal detoxification causing relevant damage to the ovarian proteome might be one of the mechanisms underlying reproductive aging. Further investigation of the role for the AGE-RAGE axis in the ovarian follicular environment is needed, and results could relate to assisted reproduction technology outcomes and new measures of ovarian reserve.

Kisspeptin signalling in the hypothalamic arcuate nucleus regulates GnRH pulse generator frequency in the rat.

BackgroundKisspeptin and its G protein-coupled receptor (GPR) 54 are essential for activation of the hypothalamo-pituitary-gonadal axis. In the rat, the kisspeptin neurons critical for gonadotropin secretion are located in the hypothalamic arcuate (ARC) and anteroventral periventricular (AVPV) nuclei. As the ARC is known to be the site of the gonadotropin-releasing hormone (GnRH) pulse generator we explored whether kisspeptin-GPR54 signalling in the ARC regulates GnRH pulses.Methodology/Principal FindingsWe examined the effects of kisspeptin-10 or a selective kisspeptin antagonist administration intra-ARC or intra-medial preoptic area (mPOA), (which includes the AVPV), on pulsatile luteinizing hormone (LH) secretion in the rat. Ovariectomized rats with subcutaneous 17β-estradiol capsules were chronically implanted with bilateral intra-ARC or intra-mPOA cannulae, or intra-cerebroventricular (icv) cannulae and intravenous catheters. Blood samples were collected every 5 min for 5–8 h for LH measurement. After 2 h of control blood sampling, kisspeptin-10 or kisspeptin antagonist was administered via pre-implanted cannulae. Intranuclear administration of kisspeptin-10 resulted in a dose-dependent increase in circulating levels of LH lasting approximately 1 h, before recovering to a normal pulsatile pattern of circulating LH. Both icv and intra-ARC administration of kisspeptin antagonist suppressed LH pulse frequency profoundly. However, intra-mPOA administration of kisspeptin antagonist did not affect pulsatile LH secretion.Conclusions/SignificanceThese data are the first to identify the arcuate nucleus as a key site for kisspeptin modulation of LH pulse frequency, supporting the notion that kisspeptin-GPR54 signalling in this region of the mediobasal hypothalamus is a critical neural component of the hypothalamic GnRH pulse generator.

Twelve-hour pair housing attenuates stress-induced hyperthermia in response to conditioned stimulus

The hypothalamic gonadotropin-releasing hormone (GnRH) pulse generator regulates intermittent GnRH discharge into the pituitary portal circulation, and thereby modulates the pattern of pulsatile secretion of luteinizing hormone (LH). Various internal and external factors first modify the GnRH pulse generator activity, which then alters the pulsatile pattern of gonadotrophin secretion, and eventually influence reproductive function. We have successfully recorded characteristic increases in neuronal activity associated with pulsatile LH secretion from the ovariectomized and cycling female goat by means of a multiple unit activity (MUA) recording technique. The unitary relationship between periodic increases in MUA volleys and LH pulses is well maintained under a variety of experimental conditions, providing evidence that these MUA volleys are the consequence of GnRH pulse generator activity. A procedure that has permitted long-term recording of electrophysiological manifestation of the GnRH pulse generator activity and its research application are described.

Rites of passage through puberty: A complex genetic ensemble

Worldwide, puberty is recognized by various cultures and celebrated as a rite of passage into adulthood. The methodical drumbeat of these religious and social ceremonies foreshadows the rhythm of reproduction that, in many ways, marks the final stage of development. Despite its social and physiological significance, including perpetuation of the species, the pathways that regulate the onset of puberty have evaded traditional physiological inquiries. No clear hormonal or metabolic trigger has been identified as a switch that activates the hypothalamic gonadotropin-releasing hormone (GnRH) pulse generator (1). Instead, geneticabnormalities that preclude puberty have provided the major insights into the pathways that are critical for the development and maturation of the reproductive axis (2, 3). Perhaps the best candidate for regulating the onset of puberty is kisspeptin, the ligand for the receptor encoded by GPR54, a gene identified as a cause of recessive hypogonadotropic hypogonadism (4, 5). The report by Pitteloud et al. (6) in this issue of PNAS identifies loss-of-function mutations in the prokineticin 2 (PROK2) gene, which encodes a secreted peptide that regulates the development and migration of the olfactory tract and GnRH neuron progenitors. The PROK2 mutations caused hypogonadotropic hypogonadism in both males and females. Using Prok2 −/− knockout mice, the GnRH neuron progenitors were shown to cross the cribriform plate but fail to migrate into and populate the hypothalamus. Interestingly, olfactory tract development in humans with PROK2 mutations was variable, resulting in both nonanosmic and anosmic forms of hypogonadotropic hypogonadism. This report highlights the role of genetics as a means to unravel complex developmental processes and helps explain why anosmia is a frequent, but variable, feature of inherited forms of hypogonadotropic hypogonadism.

Androgen Induces Production of Male Effect Pheromone in Female Goats

Previously we showed that the primer pheromone responsible for the "male effect" was produced in specific skin regions of castrated male goats by androgen treatments. In the present study, we examined whether androgen can also induce production of the male effect pheromone in female goats. Capsules containing dihydrotestosterone (DHT) or testosterone (T) were subcutaneously implanted into six ovariectomized (OVX) goats for 28 days. Small skin samples were collected from the head and rump regions, and the pheromone activity of their ether extracts was examined using a bioassay that monitors the electrophysiological manifestation of the hypothalamic gonadotropin-releasing hormone pulse generator as multiple-unit activity. Behaviors of OVX goats towards ovary-intact estrous goats were also examined before and at the end of DHT or T treatment. Before androgen treatment, neither the head nor rump skin samples in OVX goats showed pheromone activity. DHT treatment induced pheromone activity in the head skin sample of six OVX goats and in the rump skin sample of two OVX goats. Similar results were obtained by T treatment. In addition, OVX goats treated with T showed masculine-type sexual behaviors such as courtship and mounting behaviors towards the estrous goats. These results demonstrate that androgen is capable of inducing primer pheromone activity in the female and suggest that the synthesis pathway of the male effect pheromone exists in both sexes in the goat.

Discovery of the hypothalamic gonadotropin-releasing hormone pulse generator and of its physiologic significance

Shortly after a specific radioimmune assay for monkey luteinizing hormone (LH) became available in the late 1960s, we began to investigate the dynamics of its secretion in a variety of physiologic and experimental circumstances. One such early study was designed to determine whether a circadian rhythm in the plasma levels of the peptide could be detected. To this end, hourly blood samples were obtained for 24 hours from chronically ovariectomized monkeys that were restrained in primate chairs. We were surprised to find frequent, apparently random, and often large LH peaks during this period that were unrelated to the time of day or to perturbations in the environment.

Further Evidence for the Role of Glucose as a Metabolic Regulator of Hypothalamic Gonadotropin-Releasing Hormone Pulse Generator Activity in Goats

The present study examined the relative importance of blood glucose vs. free fatty acids as a metabolic signal regulating GnRH release as measured electrophysiologically by multiple-unit activity (MUA) in the arcuate nucleus/median eminence region in ovariectomized, estradiol-treated goats. MUA was recorded before, during, and after: 1) cellular glucoprivation by peripheral infusion of 2-deoxy-d-glucose (2DG; 25, 50, and 75 mg/kg.h, iv); 2) peripheral hypoglycemia in response to various doses (15-195 mU/kg.h, iv) of insulin infusion; and 3) cellular lipoprivation induced by peripheral infusion of sodium mercaptoacetate (MA; 2.4 mg/kg.h alone or combined with 25 mg/kg.h of 2DG, iv), and effects on the interval of characteristic increases in MUA (MUA volleys) were examined. Infusion of the highest dose of 2DG increased the mean interval between MUA volleys, whereas the lower doses of 2DG had no effect on volley interval. The MUA volley intervals lengthened as insulin-induced hypoglycemia became profound. There was a negative correlation between MUA volley intervals and blood glucose concentrations during insulin infusion, and coinfusion of glucose with insulin returned the MUA volley interval to a normal frequency. Infusion of MA alone or MA with 2DG did not increase MUA volley intervals. These findings demonstrate that glucose availability, but not fatty acids, regulates the GnRH pulse generator activity in the ruminant. Glucose is considered a key metabolic regulator that fine-tunes pulsatile GnRH release.

Substances derived from 4-ethyl octanoic acid account for primer pheromone activity for the "male effect" in goats.

A major constituent of the characteristic "goaty odor" 4-ethyl octanoic acid (4EOA) was previously shown to have no primer pheromone activity. This was also confirmed by our own bioassay system utilizing the recording technique of neural activity of the hypothalamic gonadotropin-releasing hormone pulse generator in goats. However, when the synthetic 4EOA solution was kept at room temperature for several months, primer pheromone activity appeared in the same solution. Headspace gas chromatography/mass spectrometry analysis revealed that there were several newly formed substances in addition to 4EOA samples with primer pheromone activity. These results suggest that 4EOA derived substance(s) but not 4EOA itself is(are) primer pheromone in goats.

Central cholecystokinin-octapeptide accelerates the activity of the hypothalamic gonadotropin-releasing hormone pulse generator in goats.

To clarify central actions of cholecystokinin-octapeptide (CCK-8) on reproduction, effects of an intracerebroventricular (i.c.v.) administration of CCK-8 on the activity of the gonadotropin-releasing hormone (GnRH) pulse generator were examined in ovariectomized (OVX) goats in the absence or presence of oestradiol. Goats were chronically fitted with recording electrodes in the mediobasal hypothalamus, and electrophysiological manifestations of the GnRH pulse generator were monitored as characteristic increases in the multiple-unit activity (MUA volleys). In OVX goats, a bolus i.c.v. injection of as little as 0.01 nmol of CCK-8 induced a MUA volley with a short latency, which resulted in a significant decrease in the post-treatment volley interval compared to that in the saline injected control. Administration of higher doses of CCK-8 (0.1 and 2 nmol) did not further accelerate the occurrence of the MUA volley, but stimulatory effects were observed for a longer period than that after the 0.01 nmol injection. When goats were treated with oestradiol, while a bolus i.c.v. injection of 0.01 nmol CCK-8 had no effect, an injection of 0.1 nmol of the peptide significantly decreased the post-treatment volley interval. On continuous i.c.v. infusion of CCK-8 at 3 nmol per 200 micro l/h for 3 h, MUA volleys with shorter intervals than those in the control were successively induced without any apparent change in basal plasma luteinizing hormone levels in OVX goats. These results demonstrate that central CCK-8 strongly accelerates the activity of the GnRH pulse generator in goats.

Regulation of gonadotropin secretion: implications for polycystic ovary syndrome.

Polycystic ovary syndrome (PCOS) is a disorder characterized by hyperandrogenism and chronic anovulation. Although the etiology of PCOS is unknown, perturbations of gonadotropin secretion are one of the hallmarks of this disorder. In normal menstrual physiology, the monotropic rise of plasma follicle-stimulating hormone (FSH) during the luteal-follicular transition is critical for follicular development and subsequent ovulation. One of the mechanisms by which FSH is differentially synthesized involves the luteal slowing of gonadotropin-releasing hormone (GnRH) pulse frequency by ovarian steroids. In PCOS, plasma leutinizing hormone (LH) is commonly increased, FSH is typically in the lower follicular range, and LH (and by inference GnRH) pulse frequency is persistently rapid at approximately one LH pulse per hour. The etiology of the neuroendocrine abnormalities in PCOS remain unclear; however, recent studies have revealed decreased sensitivity of the GnRH pulse generator to inhibition by ovarian steroids, particularly progesterone. This abnormality is reversed by the androgen receptor antagonist flutamide, suggesting that elevated androgen levels may alter the sensitivity of the hypothalamic GnRH pulse generator to steroid inhibition and lead to enhanced LH secretion. As such, women with PCOS require higher levels of progesterone to slow the frequency of GnRH pulse secretion, resulting in inadequate FSH synthesis and persistent LH stimulation of ovarian androgens. The decreased sensitivity of the GnRH pulse generator may help to explain the genesis of PCOS during puberty. In normal early puberty, sleep-entrained increases in LH stimulate ovarian steroids, which subsequently suppress LH frequency and amplitude during the subsequent day. In hyperandrogenemic girls destined to develop PCOS, this nocturnal increase in ovarian steroids may not be adequate to suppress the GnRH pulse generator, leading to a persistently rapid LH pulse frequency, impaired FSH production, and inadequate follicular development.

A possible role of neuropeptide Y as a mediator of undernutrition to the hypothalamic gonadotropin-releasing hormone pulse generator in goats.

To understand central mechanisms for nutritional infertility, the activity of the GnRH pulse generator was directly assessed in ovariectomized (OVX) goats under several experimental conditions by recording characteristic increases in the multiple-unit activity (volleys). When estradiol (E(2))-treated animals were fasted for 4-5 days, the activity of the GnRH pulse generator was gradually suppressed, and the volley interval at the end of fasting was significantly prolonged, compared with that during the feeding period (67.4 vs. 49.3 min, n = 5, P < 0.01). On the other hand, such a significant effect on the pulse generator was not observed in OVX goats. In the second experiment, the animals received a bolus intracerebroventricular injection of several doses (0, 2, 5, and 20 microg/400 microl) of neuropeptide Y (NPY). Exogenous NPY dose-dependently inhibited the pulse generator activity. At the highest dosage, the 1st posttreatment volley interval was significantly longer than that of the pretreatment (112.4 vs. 32.6 min, n = 5, P < 0.01) in OVX goats. The suppressive effect of NPY was similarly observed in OVX+E(2) goats. Further, when NPY was infused (10 microg/200 microl.h for 6 h) into OVX goats, the activity of the GnRH pulse generator was almost completely inhibited during the infusion period. Hypothalamic sites responding to fasting were immunohistochemically evaluated using an antibody for Fos in castrated goats. Fos-immunoreactive neurons were found in areas adjacent to the third ventricle. Double-labeling immunohistochemistry revealed that a subpopulation of NPY neurons in the arcuate nucleus was activated in response to fasting. These results demonstrate that: 1) the activity of the GnRH pulse generator is suppressed by fasting in the presence of E(2); 2) exogenous NPY inhibits the activity of the GnRH pulse generator regardless of the presence of E(2); and 3) several hypothalamic neurons or regions, including those containing NPY in the arcuate nucleus, are activated by fasting. Collectively, these observations suggest that NPY acts as a mediator of undernutrition to the GnRH pulse generator.

A Possible Role of Neuropeptide Y as a Mediator of Undernutrition to the Hypothalamic Gonadotropin-Releasing Hormone Pulse Generator in Goats

A Possible Role of Neuropeptide Y as a Mediator of Undernutrition to the Hypothalamic Gonadotropin-Releasing Hormone Pulse Generator in Goats