Suppression Of Luteinizing Hormone Pulses Research Articles

Norepinephrine Neurons in the Nucleus of the Solitary Tract Suppress Luteinizing Hormone Secretion in Female Mice.

Stress impairs fertility, at least in part, via inhibition of gonadotropin secretion. Luteinizing hormone (LH) is an important gonadotropin that is released in a pulsatile pattern in males and in females throughout the majority of the ovarian cycle. Several models of stress, including acute metabolic stress, suppress LH pulses via inhibition of neurons in the arcuate nucleus of the hypothalamus that coexpress kisspeptin, neurokinin B, and dynorphin (termed KNDy cells) which form the pulse generator. The mechanism for inhibition of KNDy neurons during stress, however, remains a significant outstanding question. Here, we investigated a population of catecholamine neurons in the nucleus of the solitary tract (NTS), marked by expression of the enzyme dopamine beta-hydroxylase (DBH), in female mice. First, we found that a subpopulation of DBH neurons in the NTS is activated (express c-Fos) during metabolic stress. Then, using chemogenetics, we determined that activation of these cells is sufficient to suppress LH pulses, augment corticosterone secretion, and induce sickness-like behavior. In subsequent studies, we identified evidence for suppression of KNDy cells (rather than downstream signaling pathways) and determined that the suppression of LH pulses was not dependent on the acute rise in glucocorticoids. Together these data support the hypothesis that DBH cells in the NTS are important for regulation of neuroendocrine and behavioral responses to stress.

RF01 | PMON45 Urocortin 3 Interaction with GABA and Glutamate in the Posterodorsal Subnucleus of The Medial Amygdala Mediates Suppression of GnRH Pulse Generator Frequency in Female Mice

Abstract Stress exerts a profound impact on reproductive function and disrupts pulsatile luteinizing hormone (LH) secretion. The posterodorsal subnucleus of the medial amygdala (MePD) is an upstream modulator of the hypothalamus pituitary gonadal axis. Stress alters neuronal activity within the MePD, increasing the expression of Urocortin3 (Ucn3) while enhancing the inhibitory output from the MePD to key hypothalamic reproductive centres. Designer receptor exclusively activated by designer drugs (DREADDs) inhibition of MePD Ucn3 neurons prevents psychological stress-induced suppression of LH pulses in female mice. The neurochemical pathways in the MePD responsible for regulating the GnRH pulse generator in the presence of stress remain unknown. We investigate the neurotransmission involved in the neural circuitry responsible for the suppression of GnRH pulse generator activity by MePD Ucn3 neuronal activation. Ucn3-Cre-tdTomato female ovariectomised (OVX) mice were unilaterally injected with AAV-ChR2 and implanted with optofluid cannulae targeting the MePD. Firstly, we optically stimulated MePD Ucn3 neurons with blue light at 10 Hz, 10 mW and monitored the effect on LH pulses. Next, we combined optogenetic activation of MePD Ucn3 neurons with pharmacological antagonism of GABAA or GABAB receptors with bicuculline or CGP, respectively, and observed the effect on LH pulsatility. Finally, during optical activation of MePD Ucn3 neurons we administered a combination of NMDA and AMPA receptor antagonists, AP5 and CNQX respectively, and monitored the effect on pulsatile LH secretion. Optogenetic stimulation of MePD Ucn3 neurons suppressed pulsatile LH secretion compared to controls and administration of aCSF had no effect on LH pulse interval (MePD Ucn3 activation: 36.04 ± 2.49 vs. pre-treatment control period: 17.50 ± 0.67 vs control virus: 21.67 ± 1.67 min; mean ± SEM; p<0.0001; n=12). Intra-MePD infusion of bicuculline (BIC) or CGP during optogenetic stimulation of MePD Ucn3 neurons completely blocked the suppressive effect of MePD Ucn3 neuronal activation on LH pulsatility (MePD Ucn3 activation and BIC: 20.50 ± 2.03 vs. pre-treatment control period: 17.58 ± 0.75 min; MePD Ucn3 activation and CGP: 16.30 ± 0.63 vs. pre-treatment control period: 17.50 ± 0.77 min; n=7-10). Intra-MePD infusion of AP5 + CNQX during optogenetic stimulation of MePD Ucn3 neurons similarly completely blocked the suppressive effect of MePD Ucn3 neuronal activation on LH pulsatility (MePD Ucn3 activation and AP5 + CNQX: 19.58 ± 1.42 min; n=6). Our findings show for the first time that optogenetic stimulation of MePD Ucn3 neurons inhibits GnRH pulse generator activity via GABA and glutamate signalling within the MePD. Presentation: Saturday, June 11, 2022 1:12 p.m. - 1:17 p.m., Monday, June 13, 2022 12:30 p.m. - 2:30 p.m.

Estradiol Potentiates But Is Not Essential for Prolactin-Induced Suppression of Luteinizing Hormone Pulses in Female Rats.

Hyperprolactinemia causes infertility by suppressing gonadotropin-releasing hormone (GnRH) and luteinizing hormone (LH) secretion. Because effects of prolactin (PRL) on the hypothalamus usually require estradiol (E2), we investigated the role of E2 in PRL-induced suppression of LH pulses. Ovariectomized (OVX) rats treated with oil or E2 (OVX + E2) received a subcutaneous injection of ovine PRL (oPRL) 30 minutes before serial measurement of LH in the tail blood by enzyme-linked immunosorbent assay. E2 reduced pulsatile LH secretion. oPRL at 1.5 mg/kg further reduced LH pulse frequency in OVX + E2 but had no effect in OVX rats. The higher dose of 6-mg/kg oPRL decreased LH pulse frequency in both OVX and OVX + E2 rats, whereas pulse amplitude and mean LH levels were lowered only in OVX + E2 rats. Kisspeptin immunoreactivity and Kiss1 messenger ribonucleic acid (mRNA) levels were decreased in the arcuate nucleus (ARC) of OVX + E2 rats. oPRL decreased both kisspeptin peptide and gene expression in the ARC of OVX rats but did not alter the already low levels in OVX + E2 rats. In the anteroventral periventricular nucleus, oPRL did not change kisspeptin immunoreactivity and, paradoxically, increased Kiss1 mRNA only in OVX + E2 rats. Moreover, oPRL effectively reduced Gnrh expression regardless of E2 treatment. In this study we used tail-tip blood sampling to determine the acute effect of PRL on LH pulsatility in female rats. Our findings characterize the role of E2 in the PRL modulation of hypothalamic components of the gonadal axis and LH release, demonstrating that E2 potentiates but is not essential for the suppression of pulsatile LH secretion caused by hyperprolactinemia.

SUN-476 Corticosterone-Mediated Suppression of the Reproductive Neuroendocrine Axis in Male Mice

It is well identified that psychosocial stress disturbs reproductive hormone secretion at multiple levels of the reproductive axis. The neural and hormonal mechanisms underlying this reproductive suppression and the role of reproductive brain neuropeptides in this process is not fully understood. We previously showed that in gonadectomized (GDX) male mice, a one-time acute restraint stress strongly suppressed in vivo luteinizing hormone (LH) pulsatility within 12-18 min of stress initiation. Furthermore, similarly stressed GDX mice showed a rise in circulating corticosterone (CORT) levels by 15 min. These correlative data suggest that CORT may be an important signaling factor involved in the stress-induced suppression of LH pulses. Thus, the objective of the current work is to identify the role of CORT in mediating stress inhibition of the reproductive neuroendocrine axis. First, we performed a dose-response and time course study to determine an effective exogenous CORT treatment paradigm to mimic endogenous CORT levels during psychosocial stress. Next, to assess CORT effects on LH pulses, GDX male mice were pulse bled every 6 min for 90 min, then injected with either an oil vehicle or 1 mg/kg CORT, and pulse bled every 6 min for another 90 min. CORT-injected mice showed a reduction in LH pulses, though not as robust or for as long as in restraint stressed mice. These data suggest that while CORT is sufficient to decrease LH pulsatile secretion, additional factors may also be involved to achieve maximal stress-induced suppression. To determine if upstream neuroendocrine signaling of stimulatory (kisspeptin) and inhibitory (RFRP-3) neurons to GnRH neurons are involved in this CORT-mediated suppression, in situ hybridization was conducted in brains of vehicle- and CORT-injected mice to determine changes in Kiss1 and Rfrp mRNA content and neuronal activation. These data are currently being analyzed. Our findings suggest that while acute CORT is capable of moderately suppressing reproductive hormone secretion, it is not the only mechanism involved during psychosocial stress exposure. Future studies will need to investigate additional factors that are important.

Central Injection of Ketone Body Suppresses Luteinizing Hormone Release via the Catecholaminergic Pathway in Female Rats

Ketosis is found in various pathophysiological conditions, including diabetes and starvation, that are accompanied by suppression of gonadal activity. The aim of the present study was to determine the role of ketone body in the brain in regulating pulsatile luteinizing hormone (LH) secretion in female rats. Injection of 3-hydroxybutyrate (3HB), a ketone body, into the fourth cerebroventricle (4V) induced suppression of pulsatile LH secretion in a dose-dependent manner in ovariectomized (OVX) rats with an estradiol (E2) implant producing diestrus plasma E2 levels. Plasma glucose and corticosterone levels increased immediately after the 4V 3HB injection, suggesting that the treatment caused a hunger response. The 3HB-induced suppression of LH pulses might be mediated by noradrenergic inputs to the hypothalamic paraventricular nucleus (PVN) because a local injection of α-methyl- p-tyrosine, a catecholamine synthesis inhibitor, into the PVN blocked 3HB-induced suppression of LH pulses and PVN noradrenaline release was increased by 4V 3HB injection in E2-primed OVX rats. These results suggest that ketone body sensed by a central energy sensor in the hindbrain may suppress gonadotropin release via noradrenergic inputs to the PVN under ketosis.

A role for the medial preoptic area in CGRP-induced suppression of pulsatile LH secretion in the female rat

Calcitonin gene-related peptide (CGRP) is involved in a variety of stress responses and plays a pivotal role in stress-induced suppression of the GnRH pulse generator in the rat. Intracerebroventricular administration of CGRP suppresses luteinizing hormone (LH) pulses and increases Fos expression within the medial preoptic area (mPOA) and paraventricular nucleus (PVN). The aims of the present study were to investigate whether the mPOA or PVN are sites of action for CGRP-induced suppression of LH pulses and whether lipopolysaccharide (LPS), restraint or insulin-induced hypoglycaemia, stressors known to suppress LH pulses, affect mRNA expression for CGRP and its receptor subunits (calcitonin receptor-like receptor (CL) and RAMP-1) in the mPOA and PVN. Micro-infusion of CGRP (50, 250 or 500 pmol) into the mPOA, but not the PVN, dose-dependently suppressed LH pulse frequency. LPS, restraint and hypoglycaemia suppressed RAMP-1 mRNA, but not CL or CGRP mRNA expression in the mPOA. In the PVN, all three stressors suppressed CL mRNA expression, but only LPS or restraint suppressed RAMP-1 mRNA, and CGRP mRNA was unaffected. These results provide evidence that, unlike the PVN, the mPOA might play an important role in the inhibitory effect of CGRP on pulsatile LH secretion. Additionally, CGRP receptor function may be involved in this brain region in stress-induced suppression of the GnRH pulse generator.

Resumption of Ovarian Cyclicity in Post‐partum Beef and Dairy Cows

There is a variable anoestrous period following parturition in the cow. Follicular growth generally resumes within 7-10 days in the majority of cows associated with a transient follicle-stimulating hormone (FSH) rise that occurs within 3-5 days of parturition. Dairy cows that are not nutritionally stressed generally ovulate their first post-partum dominant follicle (approximately 15 days), whereas beef suckler cows in good body condition normally have a mean of 3.2 +/- 0.2 dominant follicles (approximately 30 days) to first ovulation; and beef cows in poor body condition have a mean of 10.6 +/- 1.2 dominant follicles (approximately 70-100 days) to first ovulation. The lack of ovulation of dominant follicles during the post-partum period is associated with infrequent luteinizing hormone (LH) pulses, with both suckling and low level of nutrition being implicated in the prolonged suppression of LH pulses in the absence of progesterone. In dairy cows, the normal pattern of early resumption of ovulation may be delayed in high-yielding Holstein-type cows generally because of the effects of severe negative energy balance, dystocia, retained placental membranes and uterine infections. First ovulation in both dairy and beef cows is generally silent (i.e., no behavioural oestrus) and is generally (>70%) followed by a short cycle. The key to optimizing resumption of ovulation in both beef and dairy cows is appropriate pre-calving nutrition and management so that cows calve down in optimal body condition (body condition score; BCS; 2.75-3.0) with post-partum body condition loss restricted to <0.5 BCS units.

Central Lipoprivation-Induced Suppression of Luteinizing Hormone Pulses Is Mediated by Paraventricular Catecholaminergic Inputs in Female Rats

The present study aims to clarify the role of fatty acids in regulating pulsatile LH secretion in rats. To produce an acute central lipoprivic condition, mercaptoacetate (MA), an inhibitor of fatty acids oxidation, was administered into the fourth cerebroventricle (4V) in ad libitum fed ovariectomized (OVX) rats (0.4, 2, and 10 micromol/rat) with or without an estradiol (E2) implant producing diestrus plasma E2 levels. Pulsatile LH secretion was suppressed by 4V MA administration in a dose-dependent manner in both OVX and OVX plus E2 rats. Mean LH levels and LH pulse frequency and amplitude were significantly reduced by the highest dose of MA in OVX rats, and by the middle and highest dose of MA in E2-treated rats, suggesting that estrogen enhanced LH suppression. Blood glucose levels increased immediately after the highest dose of MA in both groups. Fourth ventricular injection of trimetazidine (2 and 3 micromol/rat), another inhibitor of fatty acids oxidation, also inhibited pulsatile LH release, resulting in significant and dose-dependent suppression of LH pulse frequency and an increase in blood glucose levels in OVX plus E2 rats. In contrast, peripheral injection of the highest 4V dose of MA (10 micromol/rat) did not alter LH release or blood glucose levels. Microdialysis of the hypothalamic paraventricular nucleus (PVN) revealed that norepinephrine release in the region was increased by 4V MA administration. Preinjection of alpha-methyl-p-tyrosine, a catecholamine synthesis inhibitor, into the PVN completely blocked the lipoprivic inhibition of LH and the counter-regulatory increase in blood glucose levels in OVX plus E2 rats. Together, these studies indicate that fatty acid availability may be sensed by a central detector, located in the lower brainstem to maintain reproduction, and that noradrenergic inputs to the PVN mediate this lipoprivic-induced suppression of LH release.

Paraventricular .ALPHA.1- and .ALPHA.2-Adrenergic Receptors Mediate Hindbrain Lipoprivation-induced Suppression of Luteinizing Hormone Pulses in Female Rats

Acute central lipoprivation suppresses pulsatile luteinizing hormone (LH) release and increases blood glucose levels through noradrenergic input to the hypothalamic paraventricular nucleus (PVN) in female rats. The present study was conducted to identify adrenergic receptor subtypes involved in central lipoprivation-induced suppression of pulsatile LH secretion and increases in plasma glucose levels in female rats. Acute hindbrain lipoprivation was produced by injection into the fourth cerebroventricle (4V) of 2-mercaptoacetate (MA), an inhibitor of fatty acid oxidation, in estradiol-implanted ovariectomized rats. Two min before MA injection, alpha1-, alpha2- or beta-adrenergic receptor antagonist was injected into the PVN. Injection of MA into the 4V suppresses pulsatile LH release in PVN vehicle-treated rats, whereas pretreatment of animals with injection of alpha1- or alpha2-adrenergic antagonist into the PVN blocked the effect of the 4V MA injection on LH pulses. beta-Adrenergic antagonist did not affect MA-induced suppression of LH pulses. The counter-regulatory increase in plasma glucose levels after 4V MA injection was also partially blocked by pretreatment with alpha1- and alpha2-adrenergic receptor antagonists. These results suggest that alpha1- and alpha2-adrenergic receptors in the PVN mediate hindbrain lipoprivation-induced suppression of LH release and counter-regulatory increases in plasma glucose levels in female rats.

MOLECULAR SPECIES OF PROSTAGLANDINS MEDIATING STRESS-INDUCED SUPPRESSION OF GONADOTROPIN SECRETION IN THE BRAIN

It is well established that reproductive function is suppressed under stress conditions. We have previously shown that all the infectious (lipopolysaccharide), hypoglycemic (2-deoxyglucose) and restraint stress suppressed pulsatile release of luteinizing hormone (LH), which was associated with an increase in cyclooxygenase-2 expression in the brain. The suppression of LH pulses by these stressors was much more enhanced by andrenalectomy. On the other hand, both corticosterone and indomethacin similarly attenuated the suppressive effect of the stressors on LH pulses. These observations suggest that prostaglandins (PG) are involved in stress-induced suppression of LH pulses. In the present study, in order to determine which subtype of PGs is involved in suppressing LH pulses, we intracerebroventricularly administered 3 major PG subtypes, i.e. PGF2α, PGD2, and PGE2. We observed LH pulses in ovariectomized rats for 3 hours by sequential blood sampling of 5 min-intervals. PGs were injected 1 h after the start of blood sampling. First, we treated animals with each PG in 3 increasing doses (0.1, 1 and 10 μg). PGF2α did not have any influence on LH pulsatility at all the doses used. PGD2 induced a dose-dependent increase in the amplitude of LH pulses while it did not affect the frequency of LH pulses and mean LH concentrations. On the other hand, PGE2 dose-dependently induced a transient increase and subsequent decreased in both mean LH concentrations and pulse amplitude, and strongly suppressed LH pulse frequency at the highest dose. Secondly, we co-injected PGF2α (5 μg) and PGE2 (5 μg), whose syntheses are known to simultaneously increase under infectious stress conditions. This resulted in a transient suppression of LH pulses in all the 3 parameters; frequency, amplitude, and mean concentrations. These results, together with our previous findings, suggest that PGE2, independently or together with PGF2α, in the brain plays a role in suppressing LH pulses as a common mediator of stressors, and that glucocorticoids maintain reproductive function by decreasing the synthesis of these PGs. (poster)

Neuroendocrine mechanism mediating energetic regulation of gonadotropin release in female rats

ABSTRACTEnergy level is a critical factor controlling gonadal activity at various phases of reproduction. A female rat model has revealed that fasting‐induced luteinizing hormone (LH) suppression is mediated by a specific neural pathway, such as noradrenergic neurons originating in the A2 region and projecting to the hypothalamic paraventricular nucleus and corticotropin‐releasing hormone neurons. The pathway is shared with that mediating glucoprivic suppression of LH pulses. Among the peripheral signals altered by energy deficiency, glucose could be a signal molecule conveying the peripheral information to the brain to regulate feeding and gonadotropin‐releasing hormone/LH release through the noradrenergic pathway during undernutrition. The brain detects the energy availability to control feeding and reproductive function at various phases of an animal’s life. It is most likely that the central glucose‐sensing mechanism could be similar to the pancreatic one, involving a glucokinase‐mediated process to detect glucose availability. Further studies are needed to elucidate the mechanism integrating the energy signals.

Acute Lipoprivation Suppresses Pulsatile Luteinizing Hormone Secretion without Affecting Food Intake in Female Rats

The present study examined the effect of acute lipoprivation on pulsatile luteinizing hormone (LH) secretion in both normal-fat diet, ad libitum-fed and fasted female rats. To produce an acute lipoprivic condition, mercaptoacetate (MA), an inhibitor of fatty acid oxidation, was administered intraperitoneally to ad libitum-fed or 24-h fasted ovariectomized (OVX) rats with or without an estradiol (E2) implant, that produces a negative feedback effect on LH pulses. The steroid treatment was performed to determine the effect of estrogen on lipoprivic changes in LH release, because estrogen enhances fasting- or glucoprivation-induced suppression of LH pulses. Pulsatile LH secretion was suppressed by MA administration in a dose-dependent manner in the ad libitum-fed OVX and OVX+E2 rats. LH pulses were more severely suppressed in the 24-h-fasted OVX and OVX+E2 rats compared to the ad libitum-fed rats. Estrogen slightly enhanced lipoprivic suppression but the effect was not significant. In the present study, increased plasma glucose and free-fatty acid concentrations may indicate a blockade of fatty acid metabolism by the MA treatment, but food intake was not affected by any of the MA doses. Acute vagotomy did not block lipoprivic suppression of LH pulses. Thus, the present study indicates that lipid metabolism is important for maintenance of normal reproductive function even in rats fed a normal-fat diet and lipoprivation may be more critical in fasted animals that probably rely more heavily on fatty acid oxidation to maintain appropriate metabolic fuel levels. In addition, failure of blockade of lipoprivic LH inhibition by vagotomy suggests that lipoprivic information resulting in LH suppression is not transmitted to the brain via the vagus nerve.

Calcitonin gene-related peptide-induced suppression of luteinizing hormone pulses in the rat: the role of endogenous opioid peptides.

Calcitonin gene-related peptide (CGRP) is involved in a variety of stress responses in the rat. Central administration of CGRP activates the hypothalamo-pituitary-adrenal axis resulting in increased corticosterone secretion. We have previously shown that central CGRP suppresses the gonadotrophin-releasing hormone (GnRH) pulse generator, specifically LH pulses. Endogenous opioid peptides (EOPs) have been shown to play an important role in stress-induced suppression of the reproductive axis. The aim of the present study was to test the hypothesis that EOPs mediate CGRP-induced suppression of pulsatile LH secretion. Ovariectomized rats were implanted with intracerebroventricular (i.c.v.) and i.v. cannulae. Intravenous administration of the opioid antagonist naloxone (250 microg) completely blocked the suppression of LH pulses induced by 1.5 microg i.c.v. CGRP and significantly attenuated the suppression of pulsatile LH secretion induced by 5 microg i.c.v. CGRP. Furthermore, intravenous administration of naloxone was found to immediately restore normal LH pulse frequency in animals treated 90 min earlier with 1.5 microg i.c.v. CGRP. Co-administration (i.c.v.) of CGRP (1.5 microg) with the mu and kappa opioid receptor-specific antagonists naloxone (10 microg) and norbinaltorphimine (5 microg), respectively, blocked the CGRP-induced suppression of LH pulses, whilst i.c.v. co-administration of CGRP (1.5 microg) with the delta opioid receptor-specific antagonist naltrindole (5 microg) did not. These data provide evidence that EOPs play a pivotal role in mediating the inhibitory effects of CGRP on pulsatile LH secretion in the rat. They also suggest that the mu and kappa, but not the delta, opioid receptors may be responsible for mediating the effects of CGRP on LH pulses.

Ovarian and hormonal responses to a progesterone-releasing controlled internal drug releasing treatment in dietary-restricted goats

An experiment was conducted to evaluate the effects of dietary restriction on ovarian, endocrine (ovarian steroids and luteinizing hormone (LH) pulse) and metabolic (glucose, insulin and non-esterified fatty acid (NEFA)) profiles in goats treated with a progesterone-releasing controlled internal drug releasing (CIDR-G) device. Cycling goats were offered either a maintenance or a restricted (30% of requirement; n=4 per treatment) level of feeding. The dietary restriction was started on the day following ovulation. At 30–32 days after the start of food restriction, the goats received a prostaglandin F 2α (2 mg of dinoprost) injection followed by 10 days of CIDR-G treatment. Ovarian ultrasonographic images were monitored daily throughout the experiment and blood samples were collected daily just before the morning feeding for analysis of endocrine and metabolic profiles. Frequent blood samples (1 ml) were also collected at 10 min intervals for 8 h from −8 h to CIDR-G removal, and from 32 to 40 h after CIDR-G removal for analysis of LH pulses. Body weight was significantly ( P<0.05) decreased in the food-restricted animals. Oestrous behaviour and ovulation followed by a rise of plasma progesterone concentration were observed after the CIDR-G removal in all control animals but not in any of the food-restricted animals within 12 days after CIDR-G removal. The LH pulse frequency from 32 to 40 h after the CIDR-G removal was significantly ( P<0.05) lower in the food-restricted animals than in control animals (1.5±0.6 versus 3.8±0.5 pulses for 8 h). There was no significant difference in the glucose concentration in weekly plasma samples between control and food-restricted animals. Insulin concentrations from 2 weeks after the start of feed restriction were significantly ( P<0.05) lower in restricted animals than in control animals. The NEFA concentration in restricted animals was significantly ( P<0.05) increased after the start of feed restriction, and then decreased gradually to the basal level. The present results suggest that nutritionally induced anovulation after CIDR-G treatment is associated with a reduction in the frequency of LH pulses, and that insulin and NEFA, rather than the glucose concentration in the circulation, may be associated with the metabolic suppression of LH pulses.

The effect of oestradiol and progesterone on hypoglycaemic stress-induced suppression of pulsatile luteinizing hormone release and on corticotropin-releasing hormone mRNA expression in the rat.

Corticotropin-releasing hormone (CRH) is implicated in the suppression of pulsatile luteinizing hormone (LH) secretion by a variety of stressful stimuli; 17beta-oestradiol (E2) has been shown to modulate this inhibitory response. The present study in ovariectomized (OVX) rats was designed to investigate the effect of E2 and progesterone (P4) on hypoglycaemic stress-induced changes in pulsatile LH secretion and on the associated changes in both central and peripheral components of the hypothalamic-pituitary-adrenal axis. E2 enhanced the hypoglycaemic stress-induced suppression of LH pulses; P4 in addition to E2 further potentiated the inhibitory response. The rise in plasma corticosterone following insulin-induced hypoglycaemia (IIH) was highest in the E2 + P4 group. Nevertheless, when such levels were achieved by administration of corticosterone, the occurrence of LH pulses was completely unaffected, irrespective of ovarian steroid milieu. E2 and E2 + P4 up-regulated basal CRH mRNA expression in the paraventricular nucleus (PVN) as measured by in situ hybridization; this signal was also increased in the medial preoptic nucleus (MPN) following E2. IIH resulted in a rise in CRH mRNA in the PVN, but not in the MPN; this rise may reflect a more significant role for the PVN in the present context. Changes in neuropeptide mRNA expression may signal changes in neuronal activity; nevertheless, the profound differences in LH pulse suppression in OVX, E2 and E2 + P4 rats following IIH were not reflected in the concurrent changes in CRH mRNA in the PVN. The results suggest that while corticosterone has no acute effect on LH pulses in the rat, the up-regulation by ovarian steroids of basal CRH mRNA in the PVN and/or MPN may contribute to the central regulation of these pulses in response to stress.

Median eminence lesions reveal separate hypothalamic control of pulsatile follicle-stimulating hormone and luteinizing hormone release.

The effects of hypothalamic lesions designed to destroy either the anterior median eminence (ME) or the posterior and mid-ME on pulsatile release of follicle-stimulating hormone (FSH) and luteinizing hormone (LH) were determined in castrated male rats. In sham-operated animals, mean plasma FSH concentrations rose to peak at 10 min after the onset of sampling, whereas LH declined to a nadir during this time. In the final sample at 120 min, the mean FSH concentrations peaked as LH decreased to its minimal value. In rats with anterior ME lesions, there was suppression of LH pulses with continuing FSH pulses in 12 of 21 rats. On the other hand, in animals with posterior to mid-ME lesions, 3 out of 21 rats had elimination of FSH pulses, whereas LH pulses were maintained. Fifteen of 42 operated rats had complete ME lesions, and pulses of both hormones were abolished. The remaining 12 rats had partial ME lesions that produced a partial block of the release of both hormones. The results support the concept of separate hypothalamic control of FSH and LH release with the axons of the putative FSH-releasing factor (FSHRF) neuronal system terminating primarily in the mid- to caudal ME, whereas those of the LHRH neuronal system terminate in the anterior and mid-median eminence. We hypothesize that pulses of FSH alone are mediated by release of the FSHRF into the hypophyseal portal vessels, whereas those of LH alone are mediated by LHRH. Pulses of both gonadotropins simultaneously may be mediated by pulses of both releasing hormones simultaneously. Alternatively, relatively large pulses of LHRH alone may account for simultaneous pulses of both gonadotropins since LHRH has intrinsic FSH-releasing activity.

Sodium pentobarbitone and the suppression of luteinizing hormone pulses in the female rat: the role of hypothermia.

The role of hypothermia in the suppression of pulsatile luteinizing hormone (LH) release by the barbiturate sodium pentobarbitone has been investigated in ovariectomized rats. Each animal was fitted with an intraperitoneal miniature radio transmitter to monitor core temperature and with an indwelling intravenous and intraperitoneal catheter. During the 6-h sampling period the animal's core temperature was recorded automatically every 5 min and a 25 microliters blood sample was obtained concurrently using an automated system. After the initial 3 h of sampling either the drug or the vehicle was administered via the intraperitoneal cannula from outside the cage, thus ensuring minimal disturbance to the animal. Administration of sodium pentobarbitone (40 mg/kg) at an ambient temperature of 21 degrees C resulted in a significant hypothermia throughout the 3-h post-injection period. During this period there was a significant reduction in mean LH concentration, and in the frequency and amplitude of the LH pulses. When the drug was administered at an ambient temperature of 35 degrees C there was no reduction in core temperature and no significant change in the LH pulse parameters. Vehicle treatment was without significant effect on core temperature or on the LH pulse parameters when administered at an ambient temperature of either 21 degrees C or 35 degrees C. These results indicate that the effects of this barbiturate on the pulsatile release of LH are secondary to the induced hypothermia and suggest that hypothermia per se may be able to disrupt LH pulses. It is therefore imperative to reassess the significance of previous studies that have implicated particular neurotransmitter systems in the control of LH pulses; unrecognized hypothermic effects of the treatments may have been the primary cause of the pulse suppression, rather than a direct involvement of the neurotransmitter in question in the regulation of LH releasing hormone. The neurophysiological process by which the hypothermic state may inhibit LH pulses remains to be determined.

Suppression of luteinizing hormone pulses by restriction of glucose availability is mediated by sensors in the brain stem

Suppression of luteinizing hormone pulses by restriction of glucose availability is mediated by sensors in the brain stem

Suppression of luteinizing hormone pulses by restriction of glucose availability is mediated by sensors in the brain stem.

The availability of metabolic fuels such as glucose is known to influence reproductive function. Peripheral administration of 2-deoxyglucose (2DG), a competitive inhibitor of glycolysis, inhibits pulsatile LH secretion in the rat and growth-retarded lamb. We hypothesized that such glucoprivic suppression of LH secretion is mediated by the lower brain stem, because studies of both ingestive and reproductive behavior implicate lower brain stem structures, such as the area postrema, as a site that is sensitive to glucose availability. In the present study, the effect of a 2DG infusion, targeted to the fourth ventricle, on pulsatile LH secretion was examined in male rats. The males were castrated or castrated and immediately implanted with testosterone. Blood samples were collected through an indwelling atrial cannula every 6 min for 4 h for LH determination. After the first hour of blood sampling, 2DG (4 or 40 mg/kg) was infused into the fourth ventricle at a flow rate of 0.2 microliter/min through a cannula that had been stereotaxically implanted 1 week before sampling. The high dose of 2DG (40 mg/kg), but not the low dose (4 mg/kg), suppressed pulsatile LH secretion and increased food intake in both castrated and testosterone-treated castrated rats. LH secretion and food intake were not affected by the infusion of xylose (40 mg/kg) as an isoosmotic control. The site specificity of the 2DG treatment was confirmed by histological examination after an isovolumetric infusion of dye (0.2 microliter/min). These results suggest that glucose availability could influence LH secretion as well as feeding through a central sensor in the lower brain stem and are consistent with the idea that the area postrema might be an important glucosensor involved in the modulation of LH secretion.

Effect of Hypothalamic Deafferentation on the Pulsatile Secretion of Luteinizing Hormone in Ovariectomized Lactating Rats

Abstract The pulsatile luteinizing hormone (LH) secretion in ovariectomized lactating rats bearing complete (CD), anterior (AD), anterolateral (ALD), posterior (PD), or roof (RD) deafferentation of the hypothalamus was determined. All lactating rats were ovariectomized on Day 2 of lactation (Day 0, day of parturition). The deafferentation of nerve fibres to the mediobasal hypothalamus was performed on Day 6 or 7 of lactation. Twenty-four h after the surgery, blood samples were taken through the indwelling atrial catheter every 6 min for 3 h. Plasma concentrations of LH and prolactin (PRL) were measured by radioimmunoassay. The loss of LH pulses associated with lactation was still apparent following AD, PD and sham-deafferentation (SD); pulsatile LH secretion was, however, present in rats with CD, ALD and RD despite continued suckling. The only significant difference in plasma PRL concentrations among the various groups was a reduction in the PRL level in rats with RD in comparison to those with SD. We conclude that the neural signal responsible for the inhibition of pulsatile LH release by suckling is conveyed through the dorsal part of the hypothalamus and PRL does not mediate the suppression of LH pulses in mid-lactation.