Rise In Plasma LH Research Articles

Gonadotropin-inhibitory hormone in Gambel's white-crowned sparrow (Zonotrichia leucophrys gambelii): cDNA identification, transcript localization and functional effects in laboratory and field experiments.

The neuropeptide control of gonadotropin secretion is primarily through the stimulatory action of the hypothalamic decapeptide, GnRH. We recently identified a novel hypothalamic dodecapeptide with a C-terminal LeuPro-Leu-Arg-Phe-NH2 sequence in the domestic bird, Japanese quail (Coturnix japonica). This novel peptide inhibited gonadotropin release in vitro from the quail anterior pituitary; thus it was named gonadotropin-inhibitory hormone (GnIH). GnIH may be an important factor regulating reproductive activity not only in domesticated birds but also in wild, seasonally breeding birds. Thus, we tested synthetic quail GnIH in seasonally breeding wild bird species. In an in vivo experiment, chicken gonadotropin-releasing hormone-I (cGnRH-I) alone or a cGnRH-I/quail GnIH cocktail was injected i.v. into non-breeding song sparrows (Melospiza melodia). Quail GnIH rapidly (within 2 min) attenuated the GnRH-induced rise in plasma LH. Furthermore, we tested the effects of quail GnIH in castrated, photostimulated Gambel's white-crowned sparrows (Zonotrichia leucophrys gambelii), using quail GnIH or saline for injection. Again, quail GnIH rapidly reduced plasma LH (within 3 min) compared with controls. To characterize fully the action of GnIH in wild birds, the identification of their endogenous GnIH is essential. Therefore, in the present study a cDNA encoding GnIH in the brain of Gambel's white-crowned sparrow was cloned by a combination of 3' and 5' rapid amplification of cDNA ends and compared with the quail GnIH cDNA previously identified. The deduced sparrow GnIH precursor consisted of 173 amino acid residues, encoding one sparrow GnIH and two sparrow GnIH-related peptides (sparrow GnIH-RP-1 and GnIH-RP-2) that included Leu-Pro-Xaa-Arg-Phe-NH2 (Xaa=Leu or Gln) at their C-termini. All these peptide sequences were flanked by a glycine C-terminal amidation signal and a single basic amino acid on each end as an endoproteolytic site. Although the homology of sparrow and quail GnIH precursors was approximately 66%, the C-terminal structures of GnIH, GnIH-RP-1 and GnIH-RP-2 were all identical in two species. In situ hybridization revealed the cellular localization of sparrow GnIH mRNA in the paraventricular nucleus (PVN) of the hypothalamus. Immunohistochemical analysis also showed that sparrow GnIH-like immunoreactive cell bodies and terminals were localized in the PVN and median eminence respectively. Thus, only the sparrow PVN expresses GnIH, which appears to be a hypothalamic inhibitory factor for LH release, as evident from our field injections of GnIH into free-living breeding white-crowned sparrows. Sparrow GnIH rapidly (within 2 min) reduced plasma LH when injected into free-living Gambel's white-crowned sparrows on their breeding grounds in northern Alaska. Taken together, our results indicate that, despite amino acid sequence differences, quail GnIH and sparrow GnIH have similar inhibitory effects on the reproductive axis in wild sparrow species. Thus, GnIH appears to be a modulator of gonadotropin release.

Effects of Passive Immunization Against Estradiol on LH and FSH in the Diestrous Female Rat.

The role of estradiol in regulating the secretion of LH and FSH from diestrus to proestrus of rats was examined by neutralizing estradiol using antiserum against estradiol (estradiol-AS). An IV injection of varying doses of estradiol-AS at 1100 h on diestrus caused a significant rise in plasma LH (maximally 318.2 ± 27.3% vs control) and non-significant rise in plasma FSH (maximally 144.0 ± 22.0% vs control) at 6 hours after the injection. Administration of the antiserum prolonged the estrous cycle and the day of ovulation by 1 day. The prolongation was caused by a 1-day delay of the LH surge. A small but significant increase in the number of tubal oocytes shed was noted in the estradiol-AS treated group (16.3 ± 0.8, n=12) compared with controls (14.5 ± 0.4, n=15). These results indicated that estradiol suppresses tonic secretion of LH and that the steroid may also be concerned with the regulation of FSH. They also confirmed that the rise in plasma estradiol from diestrous morning is necessary for triggering the LH surge on the following afternoon. Further, administration of estradiol-AS on diestrus resulted in a significant increase in the number of oocytes shed, probably due to the increase in tonic secretion of LH and FSH between diestrus to proestrus.

Acute immobilization stress alters LH and ACTH release in response to administration of N-methyl-D,L-aspartic acid in peripubertal and adult male rats.

We have evaluated the effects of stress on the pituitary hormone response to N-methyl-D,L aspartic acid (NMA) in peripubertal (43 day old) and adult (80 day old) male rats. Animals were stressed by immobilization for 3 h. Fifteen min prior to the end of the stress period, animals were injected sc with either vehicle or increasing doses of NMA (10, 30 or 60 mg/kg BW). Animals were sacrificed 15 minutes later. A significant rise in plasma LH in response to NMA occurred at a lower dose (10 versus 60 mg/kg BW), and the magnitude of the response was greater overall in peripubertal than in adult unstressed rats. Stress altered the LH response to NMA in both age groups. In peripubertal stressed rats, NMA did not induce a rise in plasma LH levels. In adult rats exposed to stress, the magnitude of the LH response to 60 mg/kg BW of NMA was lower than in unstressed animals. NMA administration triggered a rise in plasma ACTH levels in unstressed rats of both age groups, but the magnitude of the rise was greater in adults than in peripubertal animals. Stress reduced the ACTH response to NMA administration in both age groups. Plasma FSH concentrations were higher in peripubertal animals than in adults. NMA and stress were without major effect on plasma FSH levels in either age group. These data suggest that excitatory amino acids (EAA) form an important component in the regulatory processes governing the release of LH and ACTH, and that the maturational stage of the central nervous system and exposure to stress alter the LH and ACTH response to EAA.

Dopaminergic Inhibition of Gonadotropic Release in Hibernating Frogs, Rana temporaria

The influence of a dopaminergic antagonist, metoclopramide (MET), and an agonist, bromocriptine (BROMO), on reproductive status was examined in female frogs, Rana temporaria. MET induced advanced ovulation during hibernation, suggesting dopaminergic inhibition of gonadotropin (LH) release during this period. BROMO did not decrease plasma Lit in intact females in comparison with vehicle (VEH)-treated controls (VEH: 11 ± 6 vs BROMO: 5 ± 4 ng/ml) or in sham-lesioned (SL) females (SL.; 12 ± 5 vs SL + BROMO: 9 ± 8 ng/ml). However, BROMO significantly depressed the rise in plasma LH following lesioning (L) which disconnected the hypothalamus from the median eminence-pituitary complex (L + BROMO: 29 ± 10 vs L: 74 ± 30 ng/ml; P < 0.002). Taken together with previous results of lesion studies, these data point to an important role of dopaminergic inhibition in the regulation of seasonal reproduction in this frog.

Pituitary gland responsiveness to LHRH and LHRH neuronal responsiveness to excitatory stimuli are severely impaired in female rats treated neonatally with high doses of androgen

Treatment of neonatal female rats with androgen renders these animals permanently sterile as adults. Previously, we reported that these androgen-sterilized rats (ASR) do not respond to the positive feedback effects of estrogen by having LH surges. We also reported that this defect might be due to the failure of these animals to show increased hypothalamic norepinephrine turnovers (an index of secretion) in response to steroid treatment. Although LHRH-catecholamine synapses are established before or at birth, whether such synapses are functions remains to be resolved. Accordingly, the present studies, female rats were ggiven 1.25 mg of testosterone propionate at 5 days of age and, at 100 days of age, these ASR and controls were ovariectomized and treated with estradiol. In these animals, we examined whether activation of medullary A1 noradrenergic neurons would amplify LH release following preliminary depolarization of LHRH neurons with an electrochemical stimulus (ECS). As well, we reexamined whether LHRH neuronal responsiveness to exogenous NE and pituitary responsiveness to LHRH differ in controls versus ASR. In controls, two pulses of LHRH given 60 min apart elicited increases in plasma LH with the second pulse inducing greater LH release than the first pulse. In ASR, significantly less LH was released after either LHRH pulse and particularly after the second pulse. When the spacing between the two LHRH pulses was reduced to 25 min, equivalent levels of LH release occured in controls and ASR after the second pulse. The i.c.v. infusion of NE induced a rapid rise in plasma LH in both controls and ASR although the peak LH responses (at 10 min) were significantly less in the ASR group. When the medial preoptic area (MPOA) was ECS, plasma LH increased in both groups of rats but significantly less LHRH was released in ASR. Combined MPOA-ECS+i.c.v. NE in controls and ASR resulted in amplification of LH release above that obtained after only MPOA-ECS. However, the amount of LH released in ASR was significantly less than controls. In the final study, bilateral MPOA-ECS was performed and 20 min later the right A1 cell group was electrically stimulated (ES) for 15 min. In control rats, combined MPOA-ECS+A1-ES resulted in greater LH release than occured after MPOA-ECS alone. Although similar combined preoptic+A1 stimulation also amplified LH release in ASR, the plasma LH concentrations were significantly less than controls. From these studies we conclude that: (1) pituitary responsiveness to LHRH is less in ASR than controls; (2) LHRH neuronal responsiveness to various excitatory stimuli also is less in ASR compared to controls; (3) the responses of LHRH neurons to Ai-ES in controls and ASR indicate that endogenous NE signals evoke LHRH release.

Diurnal pattern of pulsatile luteinizing hormone and testosterone secretion in adult male rhesus monkeys (Macaca mulatta): influence of the timing of daily meal intake.

Adult male rhesus monkeys have a diurnal pattern of reproductive hormone secretion that is characterized by significantly elevated LH and testosterone secretion in the evening hours and a nadir in secretion of these hormones in the morning. To test the hypothesis that the daily pattern of food intake may play a role in regulating the diurnal pattern of reproductive hormone secretion we performed three studies. First, to determine the relationship between the timing of the diurnal rise in LH secretion and meal consumption, blood samples were collected from 13 adult male rhesus monkeys via chronically indwelling venous catheters (samples every 15-20 min from 0800-0800 h) while monkeys were maintained on the standard feeding regimen in our colony (one meal of Purina monkey chow fed between 1100 and 1200 h). On a day of normal feeding there was a significant diurnal rhythm in mean LH concentrations with elevated levels at night (nadir: 13.41 +/- 0.82 ng/ml from 0800-1100 h; peak: 21.34 +/- 1.56 ng/ml from 2000-2300 h, P < or = 0.05). The rising phase of the diurnal rhythm in LH secretion was apparent starting in the early afternoon, shortly after the daily meal, at 1400 h (5 h before lights went off at 1900 h), and the diurnal rise in LH secretion was no longer apparent by 0500 h (several hours before the lights went on at 0700 h). Second, we examined the influence of missing the daily meal on the diurnal pattern of LH and testosterone secretion. Blood samples were collected for a 24-h period on a day of fasting from 9 monkeys. On a day of fasting there was no diurnal rise in plasma LH or testosterone concentrations; plasma concentrations of these hormones remained at the low morning levels throughout the day. Third, we examined the diurnal pattern of LH and testosterone secretion after adapting 5 monkeys (for 6-8 weeks) to a new meal time that was 6 h later in the day than the standard meal time (i.e. at 1700 h). After adaptation to this later feeding time monkeys showed a 6-h delay in the time of the first significant rise in LH secretion (which occurred at 2000 h).(ABSTRACT TRUNCATED AT 400 WORDS)

Characterization of a Source and Levels of Plasma Immunoreactive Inhibin during the Ovulatory Cycle of the Domestic Hen1

The domestic hen with its well-organized follicular hierarchy offers a unique model system for the study of the role of inhibin in follicular development. We have validated a homologous bovine RIA for use in the hen to measure plasma immunoreactive inhibin-like activity during the ovulatory cycle as well as during experimental conditions in which follicular development has been altered. Validation of the bovine RIA kit revealed that increasing amounts of hen plasma gave linear increases in the amount of immunoreactive inhibin detected. This assay system as well as a homologous chicken LH RIA were used to assess plasma concentrations of immunoreactive inhibin and LH, respectively, in experiments 1-3. In experiment 1, blood samples were collected from hens (n = 5) at 2-h intervals for 24 h starting at 1600 h after the lay of the last egg of the sequence through the following day. No significant peak in plasma immunoreactive inhibin level was detected during the ovulatory cycle, in spite of a significant preovulatory rise in plasma LH. The highest level of plasma immunoreactive inhibin was attained, however, 2 h prior to the preovulatory LH surge. In experiment 2, eCG (n = 6) or vehicle (n = 3) was administered to hens daily for 6 days. A blood sample was drawn immediately before injection each day as well as 24 h after the last day of treatment. Treatment with eCG significantly (p < 0.01) raised plasma immunoreactive inhibin-like levels, compared to vehicle-injected controls. In experiment 3, three to four of the largest follicles (F1-F4) were surgically removed, or sham surgery was performed.(ABSTRACT TRUNCATED AT 250 WORDS)

Changes in tyrosine hydroxylase mRNA levels in medullary A1 and A2 neurons and locus coeruleus following castration and estrogen replacement in rats

Temporal changes in tyrosine hydroxylase (TH) mRNA levels in medullary A1 and A2 neurons and locus coeruleus (LC) cells were studied 6, 12 and 24 h following orchidectomy in rats. Brains from intact controls and sham castrated rats also were evaluated at these same time periods. In situ hybridization histochemistry and quantitative image analysis techniques were used to quantitate levels of cytoplasmic TH mRNA. Neither the time of day nor the stress of sham castration affected TH mRNA levels in A1, A2 and LC neurons. In contrast, 6 h following castration, TH mRNA levels in A1 neurons had declined significantly. Thereafter, there was a linear increase in A1 message levels such that, by 24 h, TH mRNA values were significantly higher than those obtained in intact controls. Placement of Silastic estrogen capsules immediately after castration prevented the 6 h decline in A1 message levels. At 12 h, TH mRNA levels in A1 neurons were significantly higher in estrogen-treated rats compared to those of the castrate or intact control groups. By 24 h, message levels in A1 neurons of steroid-treated rats were comparable to the intact control. Neither castration nor estrogen treatment altered TH mRNA levels in A2 neurons. TH mRNA levels in LC neurons increased significantly 6 h after castration and estrogen produced a further significant increase in message levels. Six hours later (12 h), TH mRNA values were still higher than controls but, in the estrogen-treated group, these levels had declined to those observed in the 12 h castrate group. By 24 h after castration, there were no differences in TH mRNA levels in any of the groups studied. Plasma LH concentrations increased significantly between 8 and 24 h and estrogen treatment completely blocked this post-castration rise. Moreover, we observed that drugs which disrupt the synthesis of NE (MPT, DDC) or which selectively block α 1-adrenoreceptors (prazosin) also prevented the post-castration rise in plasma LH. Earlier observations have shown that following castration, hypothalamic norepinephrine concentrations and turnover rates increase. The present studies suggest that, concomitant with changes in hypothalamic norepinephrine secretion, there is an increase in TH mRNA in A1 and LC neurons which may be a required preamble for the synthesis of the rate-limiting enzyme which is required to replenish the secreted norepinephrine.

Morphine but not Naloxone Enhances Luteinizing Hormone‐Releasing Hormone Neuronal Responsiveness to Norepinephrine

Some axon terminals of hypothalamic opiate neurons directly synapse on luteinizing hormone-releasing hormone (LHRH) neurons. To determine whether such synaptic connections affect LHRH neuronal activity, we have examined the profiles and concentrations of LH released in response to intracerebroventricular (icv) norepinephrine (NE, 45 μg) infusions alone or following medial preoptic area (MPOA) electrochemical stimulation (ECS) in estrogen-treated ovariectomized rats. Similar studies were performed in rats treated with naloxone (5 mg/kg ip) or morphine (20 mg/kg sc) given 15 min prior to MPOA-ECS or 30 min prior to icv NE. Naloxone neither augmented nor suppressed the LH response obtained with NE alone. MPOA-ECS evoked a significant increase in plasma LH. When the preoptic area was stimulated (0 min) and NE was infused at 30 min, a significant amplification of LH release occurred. Prior treatment of rats (-15 min) with naloxone had no effect on LH responses elicited by either preoptic stimulation alone or combined with icv NE. In the second study, morphine was given sc and had no effect on basal plasma LH levels. However, when morphine was given (-15 min) and icv NE infusions were made (30 min), the rise in plasma LH induced by NE was significantly enhanced. Preoptic ECS (0 min) evoked a rise in plasma LH and this response was also enhanced by morphine pretreatment. The major effect on LH release occurred when sc morphine injections (-15 min) were combined with MPOA-ECS (0 min) followed by icv NE (30 min). In these rats, a remarkable and highly significant release of LH occurred which reached peak levels even greater than those observed during spontaneous LH surges (2,392 versus 16 to 1,800 ng/ml). Since morphine has profound effects on the serotonergic system, in the third series of studies, morphine was infused into the dorsal raphe nucleus (DRN) and LH responses to MPOA-ECS or icv NE alone or following combined ECS + NE were examined. DRN morphine did not affect basal LH release but it produced a rapid and highly significant rise in plasma prolactin. When DRN morphine was given (-15 min) and NE was infused icv (30 min), there was marked amplification in LH release compared to those values observed after only NE. However, there were no appreciable differences in LH values obtained after sc versus DRN morphine injections in response to NE. Similarly, the amplification of LH release which occurred following DRN morphine (-15 min) + MPOA-ECS (0 min) was not different from that obtained after sc morphine. In the final group of rats, DRN morphine was given (-15 min), the preoptic area was stimulated (0 min) and NE was infused (30 min). Following this treatment, plasma LH release was also markedly enhanced and did not differ appreciably (except at 60 and 120 min) from the levels of LH released after sc morphine. Prolactin concentrations rose slowly after icv NE to reach peak levels 75 min after treatment. Combinations of morphine + MPOA-ECS without or with NE neither augmented nor suppressed the high prolactin concentrations achieved after only DRN morphine infusions. We conclude from these data that: 1) those opiate neuronal terminals which synapse directly on LHRH neurons do not affect LHRH neuronal responsiveness to either NE, to MPOA-ECS or to combined preoptic stimulation+ NE, and 2) morphine has profound effects on LHRH neuronal responsiveness to both NE, to MPOA-ECS and, in particular, to combined ECS + NE. Since amplification of LH release occurs after treatment of rats with morphine either by sc injections or DRN infusions, the augmented LH and prolactin responses observed are most likely due to the morphine-induced release of serotonin and not to direct morphine effects on LHRH neurons.

Effects of LHRH‐analogue treatment of spermatogenic dysfunction in the dog

Three male dogs, aged 2-4 years, which exhibited a high percentage of sperm with abnormal tails in their semen and low levels of peripheral plasma LH and testosterone were given two injections of 400 micrograms an LHRH analogue (LHRH-A) 3 h apart. The semen quality of all three dogs and the peripheral plasma levels of LH and testosterone were examined in two of the dogs. Plasma levels of LH and testosterone in both dogs peaked at 30 and 90 min respectively, after the injection of LHRH-A. The semen quality of all three dogs had improved temporarily 20 days after the injection. To be specific, although the mean percentage of sperm with abnormal tails in the three dogs was 19, 26 and 32% respectively before injection of LHRH-A, these values had decreased to 10, 16 and 13%, respectively, 20-40 days later. Thereafter, however, the percentage sperm abnormalities increased again. The testes and epididymides of two of the dogs were examined histologically at the end of 90 days. Sloughed germ cells were observed in the seminiferous tubules of the testes, and pyknotic epithelial cells were noted in some of the epididymal ducts. The other dog, which had a history of three unsuccessful matings was treated again with LHRH-A and mated with three bitches 30, 75, and 110 days later, respectively. Only the bitch mated with this dog 30 days after a second series of injections conceived. It is assumed that the semen quality of the three dogs improved because of a temporary rise in plasma LH and testosterone levels after the injection of LHRH-A. It is concluded that the fertility of dogs with a high percentage of sperm with abnormal tails can be improved approximately 30 days after treatment with an LHRH-A.

Thyroxine treatment prevents the development of photosensitivity in european starlings (Sturnus vulgaris)

In starlings, the breeding season is terminated by a state of photorefractoriness. Birds remain completely reproductively inactive as long as long days are maintained, and only exposure to short days restores photosensitivity. Two experiments investigated the role of different doses of thyroxine in the development of photosensitivity in castrated starlings. First, photorefractory castrated male starlings were moved from long (18L:6D) to short (8L:16D) days, and received in the drinking water either 1 or 10 mg · 1-1 thyroxine for the first 7 weeks of a 14-week observation period. Control birds regained photosensitivity after 5 weeks of short days, as signaled by a spontaneous increase in plasma LH, whereas the return to photosensitivity was delayed until weeks 7 and 9 in the 1- and 10-mg · 1-1 thyroxine-treated birds, respectively. In the second experiment, the effect of different doses of thyroxine was explored at the level of the hypothalamic Gn-RH neurosecretory neurones. The acquisition of photosensitivity in control birds transferred from long to short days was characterized by a marked increase in hypothalamic Gn-RH content (while long-day controls maintained low Gn-RH content). Doses of 10 and 20 mg · 1-1 of thyroxine completely prevented the return to photosensitivity, as seen through changes in either plasma LH concentrations or hypothalamic Gn-RH content, while a dose of 1 mg · 1-1 allowed a partial recovery of photosensitivity, as hypothalamic Gn-RH content increased to an intermediate level and the spontaneous rise in plasma LH occurred slowly but steadily.

γ-Aminobutyric Acid-A and -B Receptor Antagonists Increase Luteinizing Hormone-Releasing Hormone Neuronal Responsiveness to Intracerebroventricular Norepinephrine in Ovariectomized Estrogen-Treated Rats*

These studies determined whether endogenous gamma-aminobutyric acid (GABA) secretion affects LHRH neuronal responsiveness to norepinephrine (NE). The intracerebroventricular (icv) infusion of either bicuculline or phaclofen (GABA-A or GABA-B receptor antagonists, respectively) into ovariectomized (OVX) estrogen-treated rats did not affect basal LH levels (95 +/- 8.5 ng/ml) obtained over the 120 min of this study. When NE was infused icv, it produced a modest rise in plasma LH, which peaked within 15 min (240 +/- 25 ng/ml) and then declined toward baseline over the next 90 min. In contrast, if bicuculline was given icv at about time zero, and NE was infused (icv) 15 min later, plasma LH secretion was markedly increased and reached a peak concentration of 723 +/- 98 ng/ml within 15 min after NE treatment. Similarly, when bicuculline was infused into the medial preoptic area (MPOA), and NE was given 15 min later (icv), a peak LH level of 726 +/- 105 ng/ml was obtained within 15 min. If phaclofen was given icv at about time zero, and NE was infused 15 min later, LH rose dramatically to reach a peak concentration of 844 +/- 126 ng/ml within 15 min; a similar amplified LH response occurred when the GABA-B antagonist was infused into the MPOA and icv NE was given 15 min later. Comparisons of the LH responses obtained over the 120 min of the study suggest that icv infusions of the GABA-B receptor antagonist were more effective in sustaining peak LH secretion than the GABA-A receptor antagonist. In other groups of rats, the MPOA was electrochemically stimulated (ECS), and the effects of icv NE alone or combined with GABA receptor antagonists were evaluated. MPOA ECS alone induced a significant rise in plasma LH, which peaked between 35-45 min and then declined to approach basal levels by 150 min. In a second group, the MPOA was ECS, and at 30 min NE was infused icv. Plasma LH levels in these rats remained significantly elevated for the next 30 min before beginning their decline. In other animals, the MPOA was stimulated, and 15 min later either bicuculline or phaclofen was infused icv. Neither drug affected the patterns or concentrations of LH obtained after MPOA ECS alone. However, when rats received MPOA ECS plus either icv bicuculline or phaclofen, and these treatments were followed 15 min later by icv infusions of NE, LH secretion patterns and levels were altered significantly.(ABSTRACT TRUNCATED AT 400 WORDS)

Pituitary-testicular function is suppressed by an LHRH antagonist but not by an LHRH agonist in the marmoset monkey

The use of pituitary desensitization by an LHRH agonist (buserelin) to examine pituitary-testicular function was investigated in a New World primate. Six adult male marmoset monkeys were injected s.c. with an LHRH agonist implant (1.5 mg in a rod 0.5 cm long). Pharmacokinetics, determined by radioimmunoassay of urinary buserelin, revealed a rapid initial release of the agonist followed by a steady decline during a 200-day period. The LHRH agonist treatment resulted in a rapid initial rise in plasma LH followed by a return to mean values similar to those seen in the control samples by 7 days after implantation. Using the present protocol, no evidence of subsequent pituitary desensitization or suppression of testicular function was observed, plasma concentrations of LH and testosterone remaining within the normal range during the 200-day study period. In contrast, pituitary-testicular function was suppressed in the male marmoset after blockade of pituitary LHRH receptors by an LHRH antagonist. Five adults were treated with a single s.c. injection of the antagonist [Ac-D-Nal(2)1,D-pCl-Phe2,D-Trp3,D-Ser(Rha)6,AzGlyNH2(10)]-LHRH at a dose of 300 micrograms/kg. The LHRH antagonist induced a marked suppression of plasma LH and testosterone by 6-8 h, the low levels being maintained for 24-48 h. These results show that, whereas treatment with an LHRH antagonist can inhibit pituitary-testicular function in the male marmoset, it may be that desensitization cannot be induced by the LHRH agonist used.

A Decrease in Opioid Tone Amplifies the Luteinizing Hormone Surge in Estrogen-Treated Ovariectomized Rats: Comparisons with Progesterone Effects*

It is well known that the estrogen-induced LH surge in ovariectomized (ovx) rats is invariably far less in magnitude than the preovulatory LH surge or that induced by progesterone (P) in estrogen-primed ovx rats. Recent studies show that a decrease in hypothalamic inhibitory opioid tone by the neural clock (NC) is responsible for the induction of the preovulatory LH surge on proestrus. Therefore, we hypothesized that the diminished LH response in estrogen-treated ovx rats may be due to an inadequate reduction in opioid tone. To test this hypothesis the effects of transiently decreasing the opioid tone with an opiate receptor antagonist, naloxone (NAL), on LH secretion in estrogen-primed, short term (5 days) and long term (4 weeks) ovx rats were examined. NAL (2 mg/h) was infused iv from 1100-1400 h on day 2 in rats receiving either sc implants (two, 15 mm each) filled with 17 beta-estradiol (300 micrograms/ml in oil) or sc estradiol benzoate (EB; 10 micrograms/rat) injections at 1000 h on day 0. For comparison of NAL- and P-induced LH responses, EB-primed short and long term ovx rats received P injection (2 mg/rat, sc) instead of NAL infusion at 1100 h. Estrogen treatment alone induced a spontaneous rise in plasma LH on the afternoon of day 2, with peak LH levels ranging between 1.5-2.4 ng/ml. NAL infusion markedly enhanced the LH surge in both groups of ovx rats. In short term ovx rats NAL-induced peak LH levels (5-6 ng/ml) were less than those observed in rats receiving supplemental P treatment or that observed previously on proestrus (10-15 ng/ml). However, in long term EB-primed ovx rats, NAL infusion evoked LH surges equivalent to those observed after P injection. In addition, analysis of the episodic LH secretion pattern showed that NAL infusion accelerated the frequency and amplitude of LH discharge and significantly changed the contour of LH episodes. These results show that a transient decrease in inhibitory opioid tone before a spontaneous LH rise in estrogen-treated ovx rats can accelerate episodic LH secretion to culminate in LH surges that resemble those induced by P and the preovulatory LH surge. Therefore, these observations are in accord with the view that the NC-induced curtailment in the inhibitory opioid tone may be inadequate in estrogen-treated rats; NAL infusion and P treatment intensify decrements in inhibitory opioid tone to reinstate the preovulatory-type LH surge in these rats.

Influence of norepinephrine administration upon pituitary hormone secretion in normal men.

This study has investigated the effects of 6.2, 12.5, 25, 50 and 100 ng/kg/min/60 min of NE infused to normal men. Blood samples were obtained every 10 min, before, during and after drug administration for 3 consecutive h. Plasma levels on NE, LH, FSH, PRL, and GH were measured in all samples. The administration of 12.5 ng/kg/min over 60 min of NE induced a significant increase (p less than 0.001) in plasma NE levels (n = 5) from a mean (+/- SE) baseline of 239 +/- 14 ng/L to 706 +/- 54 ng/L which peaked and plateaued at 40 min. The calculated area under the curve was 18562 +/- 3537 ng/L/h of NE and significantly higher (p less than 0.001) than during the h before the infusion (2358 +/- 780 ng/L/h). This increase in plasma NE correlated well with the rise in plasma LH which showed a steady increase from baseline of 7.4 +/- 1.3 mIU/ml to a significant (p less than 0.05) peak of 11 +/- 1.9 mIU/ml at the end of the infusion. Furthermore, analysis of the area under the curve revealed a greater (p less than 0.05) LH release during the NE infusion (180 +/- 18 mIU/ml/h) than before the infusion (92 +/- 17 mIU/ml/h). With the exception of the studies utilizing 12.5 ng/kg/min/60 min, all other doses of NE resulted in no significant and/or consistent changes in plasma concentration of LH, FSH, GH and PRL. Thus, the direct participation of NE in the control of LH secretion in humans seems to occur in a very narrow window.

Delayed sexual maturation induced by daily melatonin administration eliminates the LH response to naloxone despite normal responsiveness to GnRH in juvenile male rats.

Daily administration of melatonin (MT) markedly delays sexual maturation in the male Wistar rat. In this study, we have evaluated pituitary responsiveness to GnRH and the level of tonic inhibition by endogenous opioids in normal juvenile male rats and in rats with delayed sexual development induced by daily afternoon MT injection (100 micrograms, s.c.) starting at 20 days of life. Plasma LH responses to repetitive intravenous GnRH administration (100 ng/100 g body weight), or to different doses of GnRH administered subcutaneously (5-100 ng/100 g body weight) were normal in MT-treated rats both at 30 and 40 days of life despite significantly lower number of pituitary GnRH receptors and decreased pituitary gonadotropin content. One naloxone (NAL) injection (2.5-5.0 mg/kg, s.c.) produced a significant increase of plasma LH in normal 40- and 55-day-old rats, which was not seen in MT-treated rats of the same age. In contrast, no increase of plasma LH was seen in 30-day-old control rats nor in MT-treated rats at this age. Pretreatment with morphine sulfate (10 mg/kg, s.c.), or with the potent Met-enkephalin analog FK 33-824 (1.0 mg/kg, s.c.) prevented the NAL-induced rise of plasma LH in control rats at day 40 of life. In all instances, plasma PRL levels were decreased after NAL both in untreated and in MT-treated rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Evidence for a negative ultrashort loop feedback mechanism operating on the luteinizing hormone-releasing hormone neuronal system.

The present studies were designed to determine whether an ultrashort loop feedback mechanism is involved in the regulation of LHRH secretion. Daily administration of a highly potent LHRH agonist (LHRH-AGO; [D-Ala6,Des-Gly10] LHRH ethylamide) immediately after orchidectomy (ORDX) significantly attenuated the rise of plasma LH from days 2 through 10 after ORDX. Concomitantly with the diminished LH rise after ORDX, a significant increase in LHRH content in the arcuate nucleus was observed in LHRH-AGO-treated rats. Measurement of LHRH levels in hypophyseal portal blood in rats 10 days after ORDX combined with daily agonist treatment revealed a significant decrease in LHRH values in portal plasma compared with those in orchidectomized controls. Arcuate nuclei-median eminence (ME) fragments obtained from ORDX rats treated in vivo with LHRH-AGO for 5 days showed a decreased basal secretion of LHRH and a diminished response to K+ stimulation compared with the release from fragments obtained from ORDX saline-treated controls. To evaluate whether a tonic LHRH inhibitory activity operates within the ME, additional experiments were performed in which ME fragments were incubated in vitro in the presence of a potent LHRH antagonist [( D-pGlu1,D-Phe2,D-Trp3,6]LHRH). The antagonist significantly enhanced the basal secretion of LHRH in a dose-dependent manner. The latter results suggest that LHRH antagonists may enhance LHRH release, perhaps by interacting with LHRH receptors playing an inhibitory role on the endogenous secretion of the decapeptide. These observations strongly suggest a tonic inhibitory or modulatory role of LHRH neurons in the regulation of their own function.

Sex difference in the effect of mating on the pulsatile secretion of luteinizing hormone in a reflex ovulator, the ferret.

Sex differences in the pulsatile secretion of LH were examined in male and female ferrets after mating. Female ferrets which were either gonadally intact and in estrus or gonadectomized and maintained on a pulsed regimen of daily estradiol (E2) injections exhibited a prolonged rise in plasma LH, characterized by an elevation in mean LH levels and an increase in the number of LH pulses after receiving an intromission from a stud male. By contrast, no such increase in LH secretion occurred in males which achieved an intromission with a female, regardless of whether they were gonadally intact and in breeding condition or gonadectomized and given pulsed estrogen. In fact, intact breeding males which achieved an intromission had significantly fewer LH pulses 1-5 h later than unmated males bled serially over the same time period. This decrease in LH pulse frequency was followed by a significant rise in mean plasma levels of androgen 5-12 h later. When a sexually dimorphic LH response to intromission was observed in gonadectomized E2-treated ferrets, we asked whether this could reflect a sex difference in pituitary responsiveness to the endogenous release of GnRH. Thus, plasma LH levels were measured in gonadectomized and gonadectomized E2-treated ferrets for 2 h after iv injection of GnRH. In the absence of gonadal steroids, ferrets exhibited a sex-specific difference in LH responsiveness to GnRH; however, no sex difference was apparent under the influence of E2. These findings demonstrate that ferrets' sexually dimorphic LH responses to intromission probably reflect a sex difference in the processing of somatosensory inputs from the genitalia or in the neural control of GnRH release into the pituitary portal vessels.

Short-term pituitary desensitization to LH-RH after pulsatile LH-RH in the ovariectomized rat: an in vivo experiment.

The development of acute insensitivity of pituitary LH secretion to LH-RH after a short exposure to LH-RH is described. In the first experiment, ovariectomized (OVX), phenobarbital-pretreated rats were given pulses of LH-RH (1.25 or 6.25 ng/100 g body weight (b.w.), intravenously). In rats given 1.25 ng at time 0, 6.25 ng at 60 min, 1.25 ng at 80 min and 1.25 ng at 120 min, there was a substantial increase in plasma LH after the first two injections, no increase after the third injection and a relatively small increase after the fourth one. In other rats treated identically but not given a 1.25-ng dose at 80 min, the plasma LH rise in response to the 1.25-ng dose at 120 min was comparable to that seen after the 1.25-ng dose given at time 0. If the 1.25-ng LH-RH pulses given at times 0 and 80 min were replaced by a rat pituitary extract, the plasma LH rise in response to the 1.25-ng dose at 120 min was comparable to that seen after administration of pituitary extract. In the second experiment, OVX phenobarbital-pretreated rats were given 1.25 ng LH-RH/100 g b.w. at t = 0. They were then divided into three groups, each receiving 1.25, 3.75 or 6.25 ng LH-RH/100 g b.w. at t = 60 min. Each of these three groups was again divided into three groups which received 1.25 ng LH-RH/100 g b.w. at 80, 100 or 120 min.(ABSTRACT TRUNCATED AT 250 WORDS)

Estrogen Changes as a Critical Factor in Modulation of Central Opioid Tonus: Possible Correlations with Post-Menopausal Migraine

The effects exerted by ovarian steroids on the modulation of opioid activity were investigated in post-menopausal migraine sufferers and in healthy controls. In order to evaluate central opioid tonus, plasma LH rise after naloxone injection was measured, bearing in mind the tonic inhibition of endogenous opioid on hypothalamic LH-RH. There was no response of plasma LH to naloxone in post-menopausal women or in patients submitted to ovariectomy in fertile life. When the subjects underwent a sequential estrogens + progestagens therapy, such a response was noted from the first month of treatment; progestagens alone were ineffective. The same phenomena were also evident in post-menopausal migraine sufferers. These data indicate that ovarian steroids modulate the activity of opiate receptors in both healthy women and migraine sufferers. Interestingly, replacement therapies through ovarian steroids restored the activity of central opioid tonus in patients affected by migraine.