Luteinizing Hormone-releasing Hormone Release Research Articles

Evidence for the presence of noradrenergic neurons and their inhibitory action on luteinizing hormone-releasing hormone release in cultured fetal rat hypothalamic cells

The control of LHRH release by catecholamine systems during fetal life (embryonic stages) was studied using hypothalamic neurons in primary cell cultures and an attempt was made to characterize the receptor type involved. Phenylephrine and clonidine, respectively α 1 an α 2 adrenoreceptor agonists, both inhibited LHRH release. These agonist inhibitory induced-effects were antagonized by the respective α 1 and α 2 adrenoreceptor antagonists (prazosin and rauwolscine). Both prazosin and rauwolscine applied alone induced a marked increase in LHRH release. Similarly, inhibition of catecholamine synthesis obtained by α-methyl-para-tyrosine (α-MT) led to a significant increase in LHRH release. The stimulatory effects induced by α 1 and α 2 adrenoreceptor antagonists or by α-MT on LHRH release suggest the presence of noradrenergic and/or adrenergic cells in fetal hypothalamic cultures. Therefore, catecholamine contents were measured in fetal hypothalamic cells in culture. Measurable amounts of norepinephrine and dopamine were found in cells, although epinephrine was undetectable. These results show: 1 - noradrenergic cells are present in primary culture of fetal hypothalamic cells. 2 - This intrinsic hypothalamic noradrenergic system exerts an inhibitory control on LHRH release at an early stage of development through α 1 and α 2 adrenoreceptors.

Estradiol modulates the LH release response to N-methyl-D-aspartate in adult female rats: studies on hypothalamic luteinizing hormone-releasing hormone and neurotransmitter release.

We investigated the effect of ovariectomy (OVX) and subsequent estradiol benzoate (EB) treatment upon the N-methyl-D-aspartate (NMDA)-induced LH secretion in adult female rats. Furthermore, the release of LHRH, norepinephrine (NE), dopamine (DA), 5-hydroxyindoleacetic acid (5-HIAA) and gamma-aminobutyric acid (GABA) from superfused hypothalamic fragments explanted from OVX and OVX-EB rats was determined. Two weeks after OVX, animals received EB (100 mg/kg) s.c., or oil vehicle (OVX-EB or OVX groups, respectively). Two days thereafter, at 09.00 h, NMDA (15 or 30 mg/kg) was given as an i.v. bolus; blood samples were drawn before and 10 min after drug administration. In OVX rats, NMDA had no significant effect on LH levels, whereas it stimulated LH release in OVX-EB animals at both doses tested (315 and 362% from basal values, p < 0.001). For hypothalamic superfusion studies OVX and OVX-EB animals were decapitated at 09.00 h, and the mediobasal hypothalami (MBH) dissected on ice. NMDA (10(-4) M) was added to the superfusion medium for a 10 min period. Basal LHRH release (OVX: 1.41 +/- 0.18; OVX-EB: 1.59 +/- 0.28 pg/10 min/MBH) was significantly (p < 0.05) enhanced by NMDA (OVX: 2.97 +/- 0.95; OVX-EB: 2.80 +/- 0.61 pg/10 min/MBH). EB treatment had no significant effect on basal or NMDA-induced LHRH output.(ABSTRACT TRUNCATED AT 250 WORDS)

α1 adrenergic regulation of estrogen-induced increases in luteinizing hormone-releasing hormone mRNA levels and release

Prazosin, an α 1 adrenergic antagonist, was used to examine the relationship between adrenergic inputs and the stimulatory effects of estrogen on LHRH mRNA and release. Bilateral cannulae were implanted just dorsal to the preoptic area (POA). Estrous cycles were monitored daily by vaginal smears. On the morning of diestrus, each rat was ovariectomized and assigned to one of three treatment groups: Control — injected with sesame oil ( n = 5); Surge — injected with estradiol benzoate (EB, 10 μg) to produce an LH surge ( n = 5); or, Surge + Prazosin — injected with EB and a prazosin-filled inner cannula was put into the POA ( n = 6). Between 4–6 pm of the following day, rats were anesthetized, decapitated, trunk blood collected, and brains were stored in liquid nitrogen. In situ hybridization was performed using a 32P end-labelled 59-mer complementary to LHRH mRNA. Reduced silver grains, proportional to LHRH mRNA content, were quantified. Treatment with estrogen alone resulted in an LH surge and a 50% increase ( P < 0.05) in numbers of cells expressing LHRH. This estrogen-induced increase and the LH surge were completely blocked ( P < 0.01) by prazosin. Prazosin also decreased ( P < 0.01) the median number of grains per cell from 81 (Surge) to 65 grains per cell (Surge + Prazosin). When the number of grains in LHRH-expressing neurons were totalled, EB increased ( P < 0.05) LHRH gene expression by 53%, and local administration of prazosin completely blocked ( P < 0.01) this increase. Since, coincident with estradiol stimulating the LH surge, it increased potential LHRH synthetic capacity (as measured by LHRH mRNA) and since administration of prazosin into POA blocked the ability of estrogen to do so, it appears that an endogenous α 1 ligand, probably norepinephrine, is a mediator of stimulatory effects of estrogen on LHRH biosynthesis as well as release.

Ethanol alters N-methyl-DL-aspartic acid-induced secretion of luteinizing hormone releasing hormone and the onset of puberty in the female rat.

In the present study, we have evaluated the effects of ethanol (ETOH) on luteinizing hormone releasing hormone (LHRH) secretion induced by N-methyl-DL-aspartic acid (NMA) in vitro and the ability of NMA to induce precocious puberty in vivo. For the in vitro experiments, the basal and NMA-stimulated release of LHRH from arcuate nucleus-median eminence (AN-ME) fragments obtained from immature female rats was measured by RIA following static incubation in medium consisting of Krebs-Ringer bicarbonate glucose buffer. Control vials contained medium only and the test vials contained medium plus ETOH doses of 30, 50, and 70 mM. These data demonstrate that ETOH did not alter basal LHRH release, but dose-dependently blocked (p < 0.01) the NMA-induced release of the peptide during anestrus, as well as first proestrus and estrus. For the in vivo experiment, 26-day-old females began receiving saline or saline-ETOH (3 g/kg) solution by gastric gavage daily at 12.30 p.m. Each day at 2.00 and 4.00 p.m., each of the animals received subcutaneous injections of a 40-mg/kg solution of NMA in saline. Other control animals received saline gastrically, as well as subcutaneously. The timing of puberty was assessed in all animals by monitoring vaginal opening (VO) and first diestrus (D1). The mean (+/- SEM) age at VO for animals in the non-ETOH group, which received NMA, was 31.7 +/- 0.40 days. Vaginal smears revealed that D1 occurred at a mean (+/- SEM) age of 33.0 +/- 0.42 days.(ABSTRACT TRUNCATED AT 250 WORDS)

Beta-endorphin regulation of LHRH release at the median eminence level: immunocytochemical and physiological evidence in hens.

We studied the effect that beta-endorphin (beta END) might have at the median eminence (ME) on luteinizing hormone (LH)-releasing hormone (LHRH) during the ovulatory cycle of domestic hens. Thus, we assessed (a) the immunocytochemical distribution of beta END and LHRH in the hen ME, (b) the temporal changes in ME and preoptic area (POA) LHRH and beta END content, in both a spontaneous and a premature C2 ovulatory model. The premature C2 ovulation occurs 6-7 h after the administration of progesterone (P4) injected 14 h before the spontaneous second (C2) ovulation of a sequence and therefore 7-8 h earlier than expected, (c) the ME in vitro release of beta END in both models, and (d) the effect of beta END and naloxone on in vitro ME-LHRH release in the two models. In the hen, beta END cell bodies are located in the periarcuate area with axons projecting to both the ME and the POA. LHRH perikarya are located in the medial POA and anterior hypothalamus and project to the ME and infundibulum through the ventrolateral hypothalamus. In the spontaneous C2 ovulatory model, both beta END and LHRH content in the ME remained unchanged during the 14 h preceding the C2 ovulation. However, POA-LHRH content was increasing at the time of the LH surge (4 h before the expected C2 ovulation) and remained elevated until the C2 ovulation occurred. In contrast, POA-beta END content was lowest at the time of the LH surge and remained low until the C2 ovulation occurred.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of gamma-aminobutyric acid-A receptor antagonist, bicuculline, on the electrical activity of luteinizing hormone-releasing hormone pulse generator in the ovariectomized rat.

The role of GABA neurons in the control of pulsatile release of LHRH was investigated by checking the effect of the GABAA receptor agonist, muscimol, and antagonist, bicuculline, on the electrical activity of the luteinizing hormone-releasing hormone (LHRH) pulse generator in the ovariectomized rat fitted with chronically implanted electrode arrays in the medial basal hypothalamus. In untreated control animals, the hypothalamic multiunit activity (MUA) exhibited, at an average of 20.5-min intervals, characteristic increases (volleys), each of which was associated with the initiation of an LH pulse. A bolus i.v. injection of muscimol (2 mg/kg) significantly increased the interval between MUA volleys and LH pulses without affecting the pulse amplitude. Continuous i.v. infusion of saline increased the interval between MUA volleys to an average of 24.1 min without affecting the LH pulse amplitudes. Bicuculline infusion (10 mg/kg/h) altered neither the interval between MUA volleys nor the pulsatile release of LH. This was further checked in the condition where presynaptic inhibition by opioid peptides on the noradrenergic system was presumably decreased by naloxone. Naloxone infusion (0.5 or 0.7 mg/kg/h) caused the MUA volleys to occur markedly frequently, an average of 13.8-min intervals. The interval during combined infusion of bicuculline with naloxone was an average of 16.2 min, suggesting that bicuculline could not decrease the interval that was set by naloxone. The results show that, although exogenous GABAA receptor agonist is capable of inhibiting the activity of LHRH pulse generator, the reduction in the endogenous GABAA receptor activity does not cause a significant effect, suggesting a minor role of inhibitory GABA neurons in the control of pulsatile release of LHRH. Further, together with the well-known fact that activation of GABAA receptor hyperpolarizes neurons postsynaptically, it is assumed that the GABAergic system, unlike the opioidergic one, is not involved in the presynaptic inhibition of the adrenergic receptor system which is probably implicated in the frequency control of LHRH pulse generator.

Nitric oxide mediates norepinephrine-induced prostaglandin E2 release from the hypothalamus.

Nitric oxide (NO), formed by conversion of arginine to citrulline and NO by NO synthase, mediates relaxation of vascular smooth muscle. NO synthase has been demonstrated by immunocytochemical methods in neurons in various parts of the central nervous system including the hypothalamus. The latter finding suggested to us that NO might play a role in controlling the release of hypothalamic peptides. We have previously shown that norepinephrine mediates the release of luteinizing hormone-releasing hormone (LHRH) from LHRH terminals in the median eminence into the hypophyseal portal veins, which transport LHRH to the anterior pituitary gland to trigger release of luteinizing hormone from gonadotrophs. LHRH release from these terminals requires increased release of prostaglandin E2 (PGE2). PGE2 activates adenylate cyclase to produce cAMP, and then cAMP induces the exocytosis of LHRH secretory granules. In view of the evidence above and because of the developing evidence for the importance of NO in the central nervous system, it occurred to us that NO might be involved in this process. Consequently, we evaluated the role of NO in the release of PGE2 from medial basal hypothalamic fragments. As previously reported, norepinephrine (10 microM) increased PGE2 release from the hypothalamic fragments. The inhibitor of NO synthase NG-monomethyl-L-arginine (NMMA, 300 microM) blocked the stimulation of PGE2 release induced by norepinephrine but had no effect on the basal release of PGE2. Sodium nitroprusside (100 microM), which liberates NO, also elevated PGE2 release from the hypothalamic fragments. This elevation was not affected by NMMA, presumably because NMMA blocks enzymatic generation of NO but does not alter NO liberated by nitroprusside. When the NO liberated by nitroprusside was inactivated by hemoglobin (2 micrograms/ml), the effect of nitroprusside on PGE2 release was completely inhibited. Neither NMMA nor hemoglobin altered the basal release of PGE2, which indicates that NO is not responsible for basal PGE2 release. Addition of L-arginine (10 microM to 1 mM), the substrate for NO synthase, had no effect on basal PGE2 production. These results indicate that NO synthase is not activated in unstimulated hypothalamic fragments in vitro. The results suggest that norepinephrine activates NO synthase leading to the production of NO, which subsequently activates cyclooxygenase and results in the production of PGE2. PGE2 then activates adenylate cyclase leading to generation of increased cAMP, which induces exocytosis of secretory granules of LHRH and other neuropeptides released by PGE2. The indication that NO is essential to norepinephrine-induced release of PGE2 from hypothalamic fragments provides insight into the mechanism of LHRH release and the results open the possibility that the importance of NO to neuronal functions may be widespread in the nervous system.

Sexual dimorphism in the effects of mating on the in vitro release of LHRH from the ferret mediobasal hypothalamus

A sexually dimorphic pattern in the secretion of luteinizing hormone (LH) has previously been shown to occur in response to mating in an induced ovulating species, the ferret, with mating augmenting the secretion of LH in females but not in males. The aim of this study was to determine whether this dimorphic pattern in the postcoital secretion of LH reflects a dimorphic effect of mating on the neural release of luteinizing hormone releasing hormone (LHRH). The effect of mating on the in vitro release of LHRH from mediobasal hypothalami (MBH) collected from breeding male and female ferrets was studied. Luteinizing hormone releasing hormone release and content were significantly reduced in tissues from estrous females sacrificed 0.25 h after mating compared to unpaired estrous females and estrous females sacrificed 1 or 2.6 h after the mating stimulus. By contrast, the release of LHRH from MBH fragments and LHRH tissue content were equivalent in breeding males that were sacrificed 0.25 h after mating and in breeding males that were left unpaired. These data suggest that the postcoital surge of LH in the female ferret is preceded by a release of LHRH that initially depletes neuronal terminals within the MBH, whereas LHRH release, like pituitary LH secretion, is minimally affected by mating in males.

Oestrogen Positive Feedback Reduces Arcuate Proopiomelanocortin Messenger Ribonucleic Acid

Oestradiol-17ß, in its positive feedback mode, as well as stimulating the release of luteinizing hormone-releasing hormone. (LHRH), also stimulates LHRH mRNA synthesis in medial preoptic neurons. Our aim was to determine whether the action of oestradiol might be mediated in part by inhibiting arcuate proopiomelanocortin (POMC) neurons which terminate on LHRH neurons. Female rats were ovariectomized at about 1000 h of dioestrus, given a subcutaneous injection of oil or oestradiol benzoate and killed at 1600 h the same day (presumptive dioestrus), presumptive pro-oestrus and oestrus and at 1000 h of presumptive pro-oestrus. In situ hybridization of brain sections was carried out with a [(35) S]UTP-labelled riboprobe containing 538 bases of a cDNA identical to exon 3 of the POMC gene, which includes the ß-endorphin coding domain. The amount of POMC mRNA in cells of the anterior arcuate nucleus, as assessed by grain counts, was significantly decreased in oestradiol benzoate- compared with oil-treated animals at 1600 h of pro-oestrus, but at none of the other times studied. This finding, together with other data, suggests that the positive feedback stimulation of LHRH mRNA synthesis by oestradiol-17ß may be due, in part, to the disinhibition of LHRH neurons consequent on inhibition of POMC mRNA biosynthesis in arcuate neurons.

Intracerebroventricular LHRH relieves behavioral deficits due to vomeronasal organ removal

Contact between the sexes in many species is known to produce hormonal changes in the male [increases in serum luteinizing hormone (LH) and/or testosterone] that can be interpreted as due to an intracerebral release of LH-releasing hormone (LHRH). In some circumstances, these hormonal changes appear to depend on an intact vomeronasal sensory system. Exogenous LHRH is also known to facilitate mating behavior in several species. We show here that LHRH delivered into the cerebral ventricles can restore some mating behavior lost when the vomeronasal organs are removed from sexually inexperienced male hamsters. The results are consistent with our working hypothesis that intracerebral LHRH release is an intermediate in the facilitation of mating behavior by vomeronasal sensory input.

Role of 5-hydroxytryptamine receptors on luteinizing-hormone-releasing hormone release in the ovariectomized, estradiol-treated rat

The present experiments examined the effect of ketanserin [5-hydroxytryptamine-2 (5-HT 2) antagonist] and 8-hydroxy-2-(di- n-propylamino) tetralin (8-OH-DPAT) (5-HT 1 agonist) on the in vitro release of luteinizing-hormone-releasing hormone (LHRH) from the medial basal hypothalamuspreoptic area-suprachiasmatic nucleus region (MBH-POA-SCN) of ovariectomized (OVX), estradiol-(E 2) treated rats using in vitro superfusion techniques. Regularly cycling female Holtzman rats (250–300 g) were maintained on a photoperiod of 0500–1900 h light at 22 ± 2° C. Rats were ovariectomized (25–30 days) and received Silastic E 2 implants (150 μg E 2/ml sesame oil) SC 48 h prior to the in vitro superfusion. Following a control period of Krebs-Ringer Phosphate (KRP) superfusion, ketanserin (5HT 2 receptor antagonist, 1 × 10 −6 M) significantly increased LHRH release ( p < 0.05). Subsequent superfusion of 5-HT (1 × 10 −8 M) significantly decreased ( p < 0.05) the effect of ketanserin on LHRH release. The 5-HT 2 antagonist Lilly 53857 (1 × 10 −6 M or 1 × 10 −5 M) did not increase LHRH release above control levels. Neither 5-HT nor quipazine had a significant effect on LHRH release at 1 × 10 −6 M. Superfusion of 8-OH-DPAT (5-HT 1 receptor agonist 1 × 10 −5 M) significantly ( p < 0.01) increased LHRH release but subsequent superfusion of 8-OH-DPAT + pindolol (mixed 5-HT 1a,1b and a β-adrenergic receptor antagonist, 1 × 10 −6 M) or pindolol alone had no effect on LHRH release. These results suggest that the 5-HT 1 receptor plays a role in LHRH release and this effect may be related to the opposing effects of postsynaptic and autoreceptors. However, the failure of Lilly 53857 to reproduce the stimulatory effect of ketanserin on LHRH release suggests that 5-HT 2 receptors in the MBH-POASCN may not modify LH release during the estrous cycle.

Hypothalamic electrical activity that relates to the pulsatile release of luteinizing hormone exhibits diurnal variation in ovariectomized rats

The multiunit activity (MUA) was recorded in the arcuate nucleus-median eminence region in freely moving ovariectomized rats maintained under controlled lighting conditions (lights on 05.00–19.00 h) over a period of 24 h. The MUA of each rat had been confirmed to exhibit characteristic increases (volleys) in association with pulses of luteinizing hormone. In 6 MUAs examined, the frequency of MUA volleys was significantly greater in the dark period than in the light period, suggesting a diurnal variation in the release of luteinizing hormone-releasing hormone in the ovariectomized rat.

Central peptidergic mechanisms controlling reproductive hormone secretion: novel methodology reveals a role for the natriuretic peptides.

A variety of neural factors can influence reproductive hormone secretion by neuromodulatory actions within the hypothalamus or neuroendocrine actions within the anterior pituitary gland. Passive immunoneutralization and antagonist administration protocols have suggested physiological roles for a number of these factors; however, both experimental approaches have severe technical limitations. We have developed novel methodology utilizing cytotoxin cell targeting with neuropeptides linked to the toxic A chain of the plant cytotoxin ricin. With this methodology we can target and destroy in vivo or in vitro cells bearing receptors for that peptide. Ricin A chain conjugated to atrial natriuretic peptide (ANP), a neuropeptide known to pharmacologically inhibit luteinizing hormone-releasing hormone (LHRH) release, was injected into the cerebroventricular system of intact, cycling rats and ovariectomized rats. Cytotoxin conjugate treatment significantly lengthened the estrous cycle. In ovariectomized rats the luteinizing hormone surge induced by steroid priming was completely inhibited. LHRH content of the median eminences of these rats was not significantly altered. These data suggest that ANP binding to clearance receptors in the hypothalamus displaces the C-type natriuretic peptide (CNP) from the shared clearance receptor, making more CNP available to inhibit LHRH release. In the absence of cells bearing the clearance receptor all available CNP binds to the ANPR-B receptor and exerts its effect via an inhibitory interneuron, since LHRH fibers are spared by this treatment.

Neuropeptide Y is a neuromodulator of pulsatile luteinizing hormone-releasing hormone release in the gonadectomized rhesus monkey.

In a previous study, we have demonstrated that infusion of neuropeptide Y (NPY) into the stalk-median eminence (S-ME) of gonadectomized rhesus monkeys stimulated LHRH in a dose-dependent manner. This finding led us to address the following questions: 1) What are the characteristics of NPY release in vivo? 2) How does NPY release relate to LHRH release? 3) Is endogenous NPY essential to pulsatile LHRH release? To answer these questions, three experiments using push-pull perfusion were performed in adult gonadectomized rhesus monkeys. Perfusate samples from the S-ME were collected at 10-min intervals for 6 to 12-h periods, and the concentrations of LHRH and NPY in perfusates were determined by RIA. In Exp I, the release pattern of NPY and LHRH in the S-ME was independently determined in a group of 11 conscious monkeys: NPY release in the S-ME was pulsatile with an interpulse interval of 44.9 +/- 3.3 min (n = 11). This interpulse interval was similar to that seen for LHRH release (43.8 +/- 1.1 min, n = 7). Exp II was designed to determine whether NPY pulses and LHRH pulses occur synchronously and to examine whether NPY release in the S-ME is correlated with circulating LH pulses. NPY and LHRH concentrations in aliquots of the same perfusate sample from the S-ME and circulating LH levels were concurrently measured in 8 monkeys sedated with Saffan. It was found that NPY pulses were temporally correlated (P less than 0.001) with LHRH pulses, which were also temporally correlated (P less than 0.001) with LH pulses. Moreover, NPY pulses were correlated (P less than 0.05) with LH pulses. NPY peaks preceded LHRH peaks by 4.5 +/- 0.6 min, LHRH peaks preceded LH peaks by 5.5 +/- 0.6 min, and NPY peaks preceded LH peaks by 9.7 +/- 0.8 min. In Exp III, the role of endogenous NPY in LHRH release was evaluated by infusing a specific antiserum to NPY into the S-ME during push-pull perfusion in 8 conscious monkeys. Infusion of a specific antiserum to NPY into the S-ME at 1:100 and 1:1000 dilutions suppressed pulsatile LHRH release significantly (P less than 0.05). Infusion of nonimmune serum as a control was without effect. These results are summarized as follows: 1) NPY release in the S-ME is pulsatile, 2) NPY pulses occur synchronously with LHRH and LH pulses, and 3) immunoneutralization of endogenous NPY in the S-ME suppresses pulsatile LHRH release.(ABSTRACT TRUNCATED AT 400 WORDS)

Seasonal changes in thyroid-gonadal interactions in the edible dormouse, Glis glis.

This study was conducted to determine changes in thyroid-gonadal interaction in the edible dormouse during the phase of the annual cycle that corresponds to the end of the breeding season (from June to September). We evaluated intra-hypothalamic luteinizing hormone-releasing hormone (LHRH) content, and plasma concentrations of luteinizing hormone (LH), testosterone, thyroid-stimulating hormone (TSH) and thyroxine (T4) in three groups of dormice: (1) controls; (2) dormice receiving sufficient T4 supplementation to maintain June levels in control animals until September, thus counteracting the seasonal reduction of T4 that normally begins in July; and (3) thyroidectomized dormice. The effect of thyroidectomy was only detectable in June, when plasma T4 concentration in the control group was maximal, and consisted of a significant decrease in plasma testosterone levels. This provides strong support for the hypothesis that thyroid function positively influences gondal function during the breeding season. The T4 supplementation resulted in a decrease in hypothalamic LHRH concentration, suggesting that an increased LHRH release led to the observed stimulated hypophyseal secretion of LH in June and September and the increased circulating testosterone levels in September. There was no detectable effect in July and August. These results show that thyroid axis activation of the hypothalamic-pituitary-gondal system is only possible during certain phases of the annual cycle, particularly evidenced here during the breeding season. They also reinforce our conclusions drawn from the thyroidectomy results. Conversely, the summer testicular regression which normally occurs after the breeding season is no longer controlled by plasma T4 levels.(ABSTRACT TRUNCATED AT 250 WORDS)

A selective sexually dimorphic response in the median eminence neuropeptide Y

Recent studies show that neuropeptide Y (NPY) is widely distributed in the hypothalamus and that it stimulates luteinizing hormone-releasing hormone (LHRH) release from the medial basal hypothalamus (MBH) of male and female rats. The neuroendorcrine factors that regulate NPY neurosecretion in two sexes are not well understood. We have previously observed that orchidectomy (orch) in male rats decreased and testosterone (T) replacement increased NPY levels selectively in the median eminence (ME), arcuate nucleus (ARC) and ventromedial nucleus (VMN) and the KCl-evoked in vitro release of NPY from the MBH was likewise decreased after orch and restored by testosterone replacement. Now we report that these NPY responses are different in female rats following removal of the ovaries. Ovariectomy (ovx) decreased NPY concentrations in the ARC and VMN and not in the ME. Estrogen replacement restored NPY concentrations in the ARC and VMN, however, in the ME NPY concentration decreased significantly. Further, the KCl-evoked NPY release in vitro from either the MBH or ME of intact and ovx rats was similar. In an additional concurrent comparative study in male and female rats, we observed that of the 6 microdissected hypothalamic sites, NPY levels were approximately 50% less in the ME of intact females than in those of intact male rats. Again, in male rats there was a significant decrease in NPY levels in the ME, ARC and VMN after orch, whereas in female rats, NPY levels in the ARC and VMN, but not in the ME decreased after ovx. When the effects on LHRH levels in the ME were evaluated, we observed that unlike the case of ME NPY, LHRH levels decrease in the ME of both sexes in response to gonadectomy. These studies show that despite the widespread distribution and common source of hypothalamic NPY in male and female rats, a sexually dimorphic NPY response is apparent, selectively, in the ME where NPY levels are lower in females than in males and gonadectomy and appropriate steroid-replacement differentially affects NPY levels and release. In view of the observation that the sexually dimorphic response was seen in the ME NPY and not in ME LHRH, and NPY is an important excitatory signal to LHRH neurons in steroid-treated rats, we speculate that the different pattern of gonadotropin secretion seen in male and female rats may be due, in part, to disparate modes of NPY neurosecretion imposed by postnatal androgenization.

Dopaminergic regulation of luteinizing hormone-releasing hormone release at the median eminence level: immunocytochemical and physiological evidence in hens.

Theoretically, the most effective inhibitory control of hypophysiotropic luteinizing hormone-releasing hormone (LHRH) release might occur through a presynaptic inhibition of LHRH neuronal terminals at the median eminence (ME) level. Since: (a) we have recently reported the existence of synaptic contacts between dopamine- and LHRH-containing processes in the ewe ME, and (b) nutritional deprivation induces an ovulatory failure in both birds and mammals, we have assessed the possibility that the anovulatory state induced by feed withdrawal (FW) in laying hens, might be caused by a dopaminergic inhibition of LHRH release at the ME level. Laying hens at the start (35 weeks old) and end (75 weeks old) of their commercial egg-laying life were killed at 0, 1, 2 and 4 days after FW. Serum luteinizing hormone (LH) and progesterone (P4), in vitro release of LHRH by isolated ME, and LHRH content in ME and preoptic area (POA) were determined by RIA. ME content of dopamine (DA) and its main metabolite 3,4-dihydroxyphenylacetic acid (DOPAC) were assessed by LCED. The distribution of LHRH and tyrosine hydroxylase (TH)-containing processes at the ME level of the hen was determined immunocytochemically. In the hen, LHRH-containing cell bodies are localized in the anterior hypothalamus and medial POA. LHRH-containing axons project toward the ME and infundibulum through the ventral-lateral hypothalamus. TH-containing perikarya are concentrated in the arcuate nucleus and in the adjacent part of the periventricular nucleus, dorsal to the arcuate. TH-containing axons converge toward the ME and descend into the infundibulum. Dense concentrations of TH- and LHRH-containing processes are located in the lateral and mediobasal portions of the external layer of the ME, providing opportunities for synaptic interactions between them. Ovulatory failure and regression of the ovary and reproductive tract occurred 2-3 days after FW at the end, but not at the beginning of the hen's commercial egg-laying life. After FW, hens at the end of their productive life had higher (p less than 0.01) tuberoinfundibular DA turnover, produced less LHRH, and had lower serum LH and P4 than hens undergoing FW at the beginning of their productive life. In addition, in vitro release of HRH from denervated ME tissue of hens undergoing FW at the end of their commercial egg-laying life was higher and was reversed in a dose-dependent fashion by DA, but not by serotonin. Thus, the ovulatory failure associated with FW in laying hens might be caused by a presynaptic inhibition of in vivo LHRH release at the level of ME hypothalamic neuronal terminals.

Estradiol enhances the action of neuropeptide Y on in vivo luteinizing hormone-releasing hormone release in the ovariectomized rhesus monkey.

The effect of estrogen on the responsiveness of the luteinizing hormone-releasing hormone (LHRH) neurosecretory system to neuropeptide Y (NPY) stimulation was examined using a push-pull perfusion method in conscious monkeys. NPY, at doses of 10(-6) to 10(-12) M, was infused into the stalk-median eminence (S-ME) of ovariectomized monkeys with or without estrogen, while perfusates were continuously collected. LHRH in perfusates was measured by RIA. The results indicate that in both estrogen-primed and unprimed monkeys, NPY infusion into the S-ME elicited LHRH release in a dose-dependent manner (p < 0.01). Moreover, estrogen priming enhanced the responsiveness of LHRH release to NPY infusion: (1) the minimum NPY dose necessary to elicit a significant LHRH response was reduced, (2) the peak LHRH response to NPY at a dose of 10(-6) to 10(-10) M was increased, and (3) the total release of LHRH in response to NPY at doses of 10(-6) to 10(-12) M was increased. These results suggest that NPY stimulates LHRH release in the S-ME in the presence or absence of estrogen and that estrogen enhances the responsiveness of the LHRH neurosecretory system to NPY stimulation in the rhesus monkey.

Glucocorticoid receptors in LHRH neurons.

Luteinizing hormone-releasing hormone (LHRH)-producing neurons constitute the final pathway for regulation of reproductive endocrine function by the central nervous system. Chronically elevated levels of glucocorticoids exert an inhibitory effect on reproductive function. Although this is thought to be mediated in part via modulation of classical transmitters and peptides which regulate LHRH synthesis and release, it is also possible that glucocorticoids may regulate LHRH neurons directly. We localized glucocorticoid receptors (GR) in LHRH neurons in the rat central nervous system using immunocytochemistry. In males and randomly cycling females 10-24% of LHRH neurons in the medial septum-diagonal bands of Broca, and preoptic regions colocalized nuclear GR. Ovariectomy increased the percentage of GR/LHRH neurons at the level of the organum vasculosum of the lamina terminalis to 34%, half of which showed both nuclear and cytoplasmic GR. Treatment with estradiol reversed this effect. We suggest that the actions of glucocorticoids on reproductive endocrine function are mediated partly through direct modulation of LHRH gene expression and/or release by activated GR. Moreover, GR in LHRH neurons may provide a mechanism by which the gonadal steroid progesterone can affect LHRH neurons directly, despite a lack of progesterone receptors in these neurons.

Effects of angiotensin II on LHRH release, as measured by in vivo microdialysis of the anterior pituitary gland of conscious female rats.

The present experiments assessed the effects of central administration of angiotensin II (Ang II) on mean levels of luteinizing hormone-releasing hormone (LHRH) in the extracellular fluid of the anterior pituitary gland, monitored by in vivo microdialysis. Ovariectomized rats were tested under two conditions: (1) nonhormone-treated where Ang II infusion inhibits luteinizing hormone (LH) release, and (2) ovarian hormone-treated where Ang II stimulates LH secretion. Animals were ovariectomized and chronic guide cannulae were implanted, one into the lateral cerebral ventricle for infusion of Ang II and one directed toward the anterior pituitary gland for the insertion of the microdialysis probe. Approximately 1 week later, the dialysis probe was inserted and cemented into place. The length of the dialysis probe transected the pituitary gland from its dorsal to ventral aspects. Dialysis samples were collected at 15-min intervals. Levels of LHRH were continuously monitored in nonhormone-treated animals, prior to and during intracerebroventricular (i.c.v.) infusion of Ang II. The dialysis probe was removed at the end of the experiment. One week later, the same animals were treated with estrogen and progesterone and dialysis of the anterior pituitary gland was performed 3 days later using a protocol identical to the first dialysis sampling session. A separate group of animals was tested to confirm the effects of lateral ventricle infusion of this dose of Ang II on LH release. There were no detectable values of LHRH in dialysis samples from non-hormone-treated animals.(ABSTRACT TRUNCATED AT 250 WORDS)