Male stimulation of luteinizing hormone surge, progesterone secretion and ovulation in spontaneously persistent-estrous, aging rats

In aging, persistently estrous (PE) female rats, there are no estrous cycles or cyclic increases in luteinizing hormone (LH) secretion, but the sexual receptivity to the male is consistently maintained. We recently reported that caging and mating with fertile males elicits an LH surge followed by ovulation in aging PE rats. The present study examined the relationship between the LH surge, the increase in progesterone (P) secretion and ovulation in PE females exposed to males, and assessed whether intromission was essential for the male-induced pre-ovulatory LH surge. PE rats were implanted with intra-atrial cannulae. Six to eight days later, these females were individually caged with a fertile male and repeatedly sampled (once every 30 or 60 min) between 1400 and 1900 h for assays of plasma LH and P. Sexual behavior of the female was recorded and correlated with the changes in plasma LH and P values. Similar experiments were also performed on cannulated PE rats with their vaginal orifice blocked with adhesive tape during the caging and sampling session. In both experiments, over 90% of the PE females displayed a high degree of lordosis response to mounting by the male, and over 60% of those sexually receptive PE females exhibited an LH surge followed by ovulation. The male-induced preovulatory LH surge occurred in PE females without actual intromission. Caging with fertile males also elicited a marked increase in plasma P concentrations in PE rats and in PE females prevented from experiencing intromission.(ABSTRACT TRUNCATED AT 250 WORDS)     

Hypothalamic alterations and reproductive aging in female rats: evidence of altered luteinizing hormone-releasing hormone neuronal function

Prior to the age-related loss of regular estrous cycles, female rats exhibit an attenuated preovulatory LH surge, a sign that reproductive decline is imminent. Numerous studies have revealed an important role for the hypothalamus in aging of the reproductive axis in this species. Because LHRH represents the primary hypothalamic signal that regulates gonadotropin release, assessments of LHRH neuronal activity can provide a window into hypothalamic function during reproductive aging. Studies of the dynamic activity of LHRH neurons during times of enhanced secretion have revealed deficits in middle-aged females. Available data are consistent with a decline in LHRH synthesis, transport, and secretion in middle-aged females during times of increased demand for LHRH output. Moreover, the alterations noted in LHRH neuronal function could account, in part, for the attenuation and eventual loss of the preovulatory LH surge with age. Elements extrinsic to LHRH neurons undoubtedly contribute to the decline in the parameters of LHRH neuronal function observed in middle-aged females. Whether alterations intrinsic to LHRH neurons also play a role in the age-associated reduction in LHRH synthesis and secretion remains to be determined. Recent examinations of hormone profiles during the perimenopausal period suggest that a potential hypothalamic contribution to aging of the reproductive axis in women warrants further examination.     

Neuroendocrine regulation of pulsatile luteinizing hormone secretion in elderly men

Leydig cell function is driven by LH, secreted in a pulsatile manner by the anterior pituitary in response to episodic discharge of hypothalamic LHRH into the pituitary portal circulation, under control of a yet to be defined neural mechanism, the "hypothalamic LHRH pulse generator". The normal aging process in elderly men is accompanied by a decline in Leydig cell function. Whereas primary testicular factors undoubtedly play an important role in the decrease of circulating (free) testosterone levels with age, recent studies demonstrated that aging also affects the central compartment of the neuroendocrine cascade. Hypothalamic alterations comprise changes in the regulation of the frequency of the LHRH pulse generator with an inappropriately low frequency relative to the prevailing androgen impregnation and opioid tone, and with an increased sensitivity to retardation of the LHRH pulse generator by androgens. As observed by some authors in basal conditions and by others after endocrine manipulations. LH pulse amplitude seems also to be reduced in elderly men as compared to young subjects. This is most probably the consequence of a reduction in the amount of LHRH released by the hypothalamus. Indeed, challenge of the gonadotropes with low, close to physiological doses of LHRH in young and elderly men reveals no alterations in pituitary responsiveness when looking at either the response for immunoreactive LH or bioactive LH. Deconvolution analysis on data obtained after low-dose LHRH suggests a markedly prolonged plasma half-life of LH in elderly men, a finding which may explain the paradoxical increase of mean LH levels in face of the reduced or unchanged frequency and amplitude of LH pulses.     

Hypothalamic luteinizing hormone-releasing hormone (LHRH) and LH secretion in aging female rats

Age-related changes in hypothalamic luteinizing hormone-releasing hormone (LHRH) and luteinizing hormone (LH) secretion were studied in young (6 months), middle-aged (12 months) and old (18 months) female rats. The LHRH levels in the mid-hypothalamic area were higher in intact middle-aged and old females than in young ones. Additionally, there was no age difference in the hypothalamic LHRH levels in male rats. In order to clarify the significance of this age-related increase in female rats, we examined the effects of progesterone treatment in estrogen-primed ovariectomized young and old rats on the LHRH levels in the median eminence (ME) and on plasma LH levels. We found phasic changes in ME-LHRH and plasma LH levels in estrogen-primed rats following progesterone treatment in rats of both ages, but the progesterone-induced change in ME-LHRH levels tended to be delayed in old rats compared with young females. This delay may correspond to the delayed onset, slow and low magnitude of plasma LH increase in old females. The ME-LHRH levels were generally higher in old rats than in young rats. Nevertheless, we found that the increase in plasma LH in response to progesterone treatment in estrogen-primed ovariectomized females was smaller in old rats than young rats. These results suggest that the LHRH secretory mechanism changes with age in female rats. Such alterations may result in the accumulation of LHRH in the mid-hypothalamic area and an increase in ME-LHRH.     

Pancreatic polypeptides affect luteinizing and growth hormone secretion in rats

We have examined the effects of third cerebroventricular (3V) injections of avian and bovine pancreatic polypeptide (APP and BPP) and the C-terminal hexapeptide amide of human PP (CHPP) on the secretion of anterior pituitary hormones in conscious ovariectomized rats. Injection of APP (2.0 micrograms; 472 pmoles) or BPP (5.0 micrograms; 1191 pmoles) decreased plasma levels of luteinizing hormone (LH) when compared to pre-injection levels in these animals or to saline-injected controls. The lower dose of BPP (0.5 micrograms; 119 pmoles) decreased plasma LH versus pre-injection levels and control animals, however, these effects diminished at later times. Plasma growth hormone (GH) also decreased following 3V injections of APP (2.0 micrograms) or BPP (5.0 micrograms). The lower dose of BPP (0.5 microgram) initially inhibited GH release, however, this effect was rapidly reversed and GH levels were significantly greater than those in controls at 60 and 120 min. Injections of BPP or APP did not alter prolactin (PRL) or thyroid stimulating hormone (TSH) secretion. Administration of 2.0 micrograms and 0.2 microgram of CHPP (2488 and 249 pmoles) produced no significant effects on plasma LH, GH, PRL or TSH. APP and BPP had no consistent effects on hormone secretion from dispersed anterior pituitary cells. The results indicate that APP and BPP exert potent central effects which inhibit LH and GH release from the pituitary gland.     

Characteristics of pulsatile luteinizing hormone secretion in middle-aged ovariectomized rats

Experiments were performed to characterize the pulsatile patterns of circulating luteinizing hormone (LH) in the middle-aged ovariectomized (OVX) rat. Frequent blood samples were taken from OVX rats at 6, 7-8, and 9-10 mo of age, and LH was measured by radioimmunoassay. Rats had been OVX either 2 wk (STO) or 10-20 wk (LTO) previously. Mean LH levels were significantly lower with increasing age, reflecting effects on both pulse amplitude and pulse frequency. Mean LH levels were higher in LTO than STO groups, reflecting primarily an increase in pulse amplitude, but there was also a small, significant decrease in pulse frequency with increased time following OVX. In a second experiment, a random selection of the rats in the STO groups was tested again 10 wk after OVX. A significantly higher number of 9- to 10-mo-old rats had pulsatile patterns at 10 wk than at 2 wk following OVX. Furthermore, mean plasma LH concentrations were higher in all three groups. We conclude that decreases in several parameters of LH secretion are seen in middle-aged OVX rats, at the time when irregularities are first seen in the estrous cycle in the intact rat.     

Pulsatile infusion of luteinizing hormone-releasing hormone: Effects on luteinizing hormone secretion and ovarian function in prepuberal gilts

Ten intact and hypophysial stalk-transected (HST), prepuberal Yorkshire gilts, 112–160 days old, were subjected to a pulsatile infusion regimen of luteinizing hormone-releasing hormone (LHRH) to investigate secretion profiles of luteinizing hormone (LH) and ovarian function. A catheter was implanted in a common carotid artery and connected to an infusion pump and recycling timer, whereas an indwelling external jugular catheter allowed collection of sequential blood samples for radioimmunoassay of LH and progesterone. In a dose response study, intracarotid injection of 5 μg LHRH induced peak LH release (5.9 ± 0.65 ng/ml; mean ± SE) within 20 min, which was greater (P < 0.001) than during the preinjection period (0.7 ± 0.65 ng/ml). After HST, 5 μg LHRH elicited LH release in only one of three prepuberal gilts. Four intact animals were infused with 5 μg LHRH (in 0.1% gel phosphate buffer saline, PBS) in 0.5-ml pulses (0.1 ml/min) at 1.5-h intervals continuously during 12 days. Daily blood samples were obtained at 20-min intervals 1 h before and 5, 10, 20, 40, 60 and 80 min after one LHRH infusion. Plasma LH release occurred in response to pulsatile LHRH infusion during the 12-day period; circulating LH during 60 min before onset of LHRH infusion was 0.7 ± 0.16 ng/ml compared with 1.3 ± 0.16 ng/ml during 60 min after onset of infusion (P < 0.001). Only one of four intact gilts ovulated, however, in response to LHRH infusion. This animal was 159 days old, and successive estrous cycles did not recur after LHRH infusion was discontinued. Puberal estrus occurred at 252 ± 7 days in these gilts and was confirmed by plasma progesterone levels. These results indicate that intracarotid infusion of 5 μg LHRH elicits LH release in the intact prepuberal gilt, but this dosage is insufficient to cause a consistent response after HST.     

Reduced pulsatile luteinizing hormone and testosterone secretion with aging in the male rat

To identify possible age-dependent changes in the feedback relationship between the brain-pituitary and testes, we examined the minute-to-minute patterns of plasma luteinizing hormone (LH) and testosterone (T) in intact, young male rats and compared these profiles to those of old animals. Young (3 mo; n = 11) and old (22 mo; n = 12) Sprague-Dawley rats were fitted with indwelling venous catheters and between 24 and 48 h later, were bled without anesthesia, by remote sampling, at 10-min intervals for 8 h. Blood samples of 400 microliter were withdrawn, and an equivalent volume of a blood replacement mixture was infused after each sample. Plasma LH and T levels in each sample were measured by radioimmunoassay (RIA). Plasma T levels in old animals failed to show the transient oscillations observed in young animals. Mean plasma T levels were 50% lower in old compared to young animals (P less than 0.001). Plasma patterns of LH in old animals, like their younger counterparts, showed statistically significant episodic increases, whose apparent pulse frequency was inappropriately low for their circulating T level (although not statistically different from the young group). Pulse amplitude in the old animals was 66% lower in the old compared to the young group (P less than 0.015). We conclude that age-associated alterations in brain mechanisms governing LH secretion underline these endocrine changes.     

Effects of pulsatile infusion of luteinizing hormone-releasing hormone on luteinizing hormone secretion and ovarian function in hypophysial stalk-transected beef heifers

Hypothalamic regulation of luteinizing hormone (LH) secretion and ovarian function were investigated in beef heifers by infusing LH-releasing hormone (LHRH) in a pulsatile manner (1 microgram/ml; 1 ml during 1 min every h) into the external jugular vein of 10 hypophysial stalk-transected (HST) animals. The heifers were HST approximately 30 mo earlier. All heifers had increased ovarian size during the LHRH infusion. The maximum ovarian size (16 +/- 2.7 cm3) was greater (P less than 0.01) than the initial ovarian size (8 +/- 1.4 cm3). Ovarian follicular growth occurred in 4 of 10 HST heifers in response to pulsatile LHRH infusion. In 2 heifers, an ovarian follicle developed to preovulatory size, but ovulation occurred in only 1 animal after the frequency of LHRH was increased (1 microgram every 20 min during 8 h). In blood samples obtained at 20-min intervals every 5th day, LH concentrations in peripheral serum remained consistently low (0.9 ng/ml) and nonepisodic in the 10 HST heifers during infusion of vehicle on the day before beginning LHRH. In 7 of 10 HST animals, episodic LH secretion occurred in response to pulsatile infusion of LHRH. In 3 of these long-term HST heifers, however, serum LH remained at basal levels and the isolated pituitary seemingly was unresponsive to pulsatile infusion of LHRH as indicated by sequential patterns of gonadotropin secretion obtained at 5-day intervals. These results indicate that pulsatile infusion of LHRH induces LH release in HST beef heifers.     

Neuroendocrine control of diapause hormone secretion in the silkworm,Bombyx mori

To clarify the control mechanism of diapause hormone (DH) secretion in the silkworm Bombyx mori a series of anatomical and pharmacological experiments were carried out. The arrangement of 'diapause' and 'non-diapause' eggs in the ovarioles of the moths was determined by the coloration method to estimate the accumulation of 3-hydroxykynurenine in the eggs. The females destined to lay non-diapause eggs (non-diapause producers) had diapause eggs in their ovaries if their subesophageal ganglions (Sg) had been surgically removed at 2days after larval-pupal ecdysis or later. In contrast when the surgical extirpation extended to the brain and the corpora cardiaca (CC)-corpora allata (CA) complex in addition to the Sg, the non-diapause producers had no diapause eggs. When the Sg was removed from the females destined to lay diapause eggs (diapause-producers), diapause eggs appeared in response to the treatment at 2days after larval-pupal ecdysis, but the appearance of diapause eggs was delayed by 2days when the brain-CC-CA complex was included among the organs removed. These observations suggested that DH is produced in Sg and transferred to the CC-CA complex, and that the secretion of DH from the complex is suppressed in non-diapause producers. The pattern of diapause and non-diapause eggs induced by the transection of the subesophageal connective in diapause and non-diapause producers suggested a regenerative and secretory capacity of the neurosecretory cells after the operation. The appearance of diapause eggs in non-diapause producers with transected protocerebrum of the brain confirmed that there was an inhibitory center in the protocerebrum. Changes in parts of the ovarioles containing diapause and non-diapause eggs with time of injection of gamma-aminobutyric acid (GABA) and picrotoxin suggested that a GABAergic inhibitory mechanism in DH secretion may be active in non-diapause producers but inactive in diapause producers throughout the pupal stage.     

Serotonin neuroleptics change patterns of preovulatory secretion of luteinizing hormone in rats

Serotonin receptor agonists or antagonists were used in this study to determine the timing and influence of serotonergic neurotransmission on phasic secretion of luteinizing hormone (LH). Daily injections of cyproheptadine (CP) or methysergide (MS), serotonin antagonists, initiated at 1600h on the day of vaginal proestrus, blocked the LH surge and ovulation. Vaginal smears remained cornified for 2–3 days. The drugs were ineffective when given at 0800h, though they terminated the LH surge prematurely when administered at 1730h. When quipazine, a serotonin receptor agonist was injected at 1400h or 2000h on proestrus, serum LH levels rose. This effect caused the LH surge to begin prematurely or to be sustained unusually long. Quipazine injected on diestrus 2 did not cause LH levels to rise, suggesting that its effect is estrogen dependent. Serotonin turnover in the hypothalamus was greater during onset of the LH surge than during its termination. When the LH surge was prolonged by exposing rats to light on proestrous evening, serotonin turnover remained high. The results of this study indicate that phasic secretion of LH on proestrus is accompanied by and may be dependent upon a period of serotonin neural activity.     

Neuroendocrine regulation of luteinizing hormone and follicle stimulating hormone: a review

Since the pioneering studies of Everett, Sawyer and Markee (1) it has been generally accepted that the central nervous system (CNS) regulates the secretion of the pituitary gonadotropins, luteinizing hormone (LH) and follicle stimulating hormone (FSH). However, great gaps still exist in our understanding of the neural mechanisms that regulate the secretion of these hormones. The purpose of this review is to provide the reader with a concise overview of this topic. Gaps, inconsistencies and future directions of this area of research are also presented.     

Testosterone-dependent effects of galanin on pituitary luteinizing hormone secretion in male rats

Galanin is a 29-amino-acid peptide that colocalizes with GnRH in hypothalamic neurons. High concentrations of galanin are present in portal vessel blood of both male and female rats, and galanin receptors are present on gonadotropes in both sexes. Results from studies of female rats indicate that galanin acts at the level of the pituitary to directly stimulate LH secretion and also to enhance GnRH-stimulated LH secretion. The effects of galanin on pituitary LH secretion in male rats are relatively uncharacterized; thus, the present in vivo study was conducted 1). to examine the ability of galanin to affect basal or GnRH-stimulated LH secretion in male rats and 2). to determine whether the effects of galanin on LH secretion in male rats are testosterone-dependent. All three doses of galanin used (1, 5, and 10 micro g/pulse) significantly enhanced GnRH-stimulated LH secretion in intact male rats. Only the highest dose of galanin directly stimulated LH secretion (without GnRH coadministration) in intact males. Galanin did not directly stimulate LH secretion or enhance GnRH-stimulated LH secretion in castrated male rats. In fact, the highest dose of galanin inhibited GnRH-stimulated LH secretion in castrated males. Upon testosterone replacement, the ability of galanin to directly stimulate LH secretion and to enhance GnRH-stimulated LH secretion was restored in castrated males. These results suggest a role for galanin in the regulation of LH release in male rats and demonstrate that testosterone upregulates the ability of the pituitary to respond to the stimulatory effects of galanin.     

Neuroendocrine control of gonadotropin secretion

Luteinizing hormone releasing hormone (LHRH), a hypothalmic peptide that is concentrated in granules of neurons, has the capacity to release gonadotropins (luteinizing hormone (LH) and follicle stimulating hormone) from the pituitary gland. LHRH has been found in hypophysial portal blood of rats, monkeys, and rabbits. Antibodies to LHRH depress plasma LH concentrations in castrated animals and evoke testicular atrophy, but passive immunization against LHRH does not block the LH surge induced by estrogen in monkeys. Estrogens, progestin, prolactin, and dopamine have marked effects on LH secretion, yet an association between these effects and altered hypophysial portal blood concentrations of LHRH is not established. In view of the paucity of evidence demonstrating such a cause and effect relationship, two alternative proposals have become tenable. One, hormones and neurotransmitters may not alter the levels of portal blood LHRH, but rather alter the frequency of pulsatile LHRH secretion. Two, hormones, such as estrogens, progesterone, and prolactin, may alter the responsiveness of the gonadotropin-secreting cells to LHRH by affecting the secretion of dopamine.     

Isolated hypothalami from aging female rats do not exhibit reduced basal or potassium-stimulated secretion of luteinizing hormone-releasing hormone.

Serum LH levels are diminished in middle-aged rats during spontaneous or steroid-induced LH surges and following ovariectomy (ovx). The compromised LH responses are presumed to reflect age-related alterations in LHRH neurosecretion. Direct measurements of LHRH output in middle-aged females are, however, limited. The present study utilizes an in vitro perifusion paradigm to assess basal and stimulated secretory capacity of LHRH neurons in isolated hypothalamic preparations from aging female rats. Individual hypothalamic fragments from middle-aged and young proestrous, ovx, and ovx, estradiol-treated females were perifused for 6 h and effluents were collected continuously at 10-min intervals. After 4 h of unstimulated output, two 10-min depolarizing pulses of KCl were administered. Although stimulated LHRH secretion was comparable in the two age groups, basal LHRH release from aging hypothalami was significantly elevated (pbasal less than 0.001). Furthermore, endocrine influences on LHRH output from aging hypothalami were less pronounced when compared to endocrine influences on LHRH output from young hypothalami, suggesting that steroidal regulation of LHRH secretion may be impaired in middle-aged females. These data demonstrate that LHRH neurons maintain the capacity to respond to a depolarizing stimulus at the time when regular estrous cycles cease and consequently suggest the importance of altered modulation of LHRH neurosecretion to the development of reproductive senescence.     

Hypothalamic deafferentation and luteinizing hormone-releasing hormone effects on secretion of luteinizing hormone in prepubertal pigs

The control of luteinizing hormone (LH) secretion was investigated in ovariectomized, prepubertal Yorkshire pigs by comparing the effects of anterior (AHD), complete (CHD), and posterior (PHD) hypothalamic deafferentation to sham-operated controls (SOC). Gilts (n = 16) were assigned randomly to treatments, fitted with an indwelling jugular catheter, and ovariectomized 2 days before deafferentation or sham-operation (Day 0). Blood for radioimmunoassay (RIA) of LH was collected sequentially at 20-min intervals for a period of 2 h before and 24, 48, 72, and 96 h after hypothalamic deafferentation or SOC. Episodic LH release after AHD or CHD was abolished (p less than 0.01), but not after PHD or SOC. Concentrations of serum LH in AHD and CHD dropped (p less than 0.01) at 24 and 48 h after surgery. Levels of LH before and after surgery in PHD and SOC were similar (p greater than 0.05). Infusion of 25 micrograms LH-releasing hormone (LHRH) i.v. at 72 and 96 h after hypothalamic deafferentation and SOC increased (p less than 0.01) serum LH to peak levels within 15 min. after infusion; LH returned to basal levels 60-80 min later. By 96 h after surgery, LH response to LH-releasing hormone (LHRH) was less in AHD and CHD as compared with the response at 72 h postinjection. Concentrations of LH in PHD and SOC were similar (p greater than 0.05) at 72 and 96 h, respectively. The results from this study clearly indicate that neural stimuli originating or traversing the neural areas rostral to the median eminence are required for secretion of LH in the pig.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of hyperprolactinemia on the control of luteinizing hormone and follicle-stimulating hormone secretion in the male rat

Experiments were conducted to determine the effects of acute hyperprolactinemia (hyperPRL) on the control of luteinizing hormone and follicle-stimulating hormone secretion in male rats. Exposure to elevated levels of prolactin from the time of castration (1 mg ovine prolactin 2 X daily) greatly attenuated the post-castration rise in LH observed 3 days after castration. By 7 days after castration, LH concentrations in the prolactin-treated animals approached the levels observed in control animals. HyperPRL had no effect on the postcastration rise in FSH. Pituitary responsiveness to gonadotropin hormone-releasing hormone (GnRH), as assessed by LH responses to an i.v. bolus of 25 ng GnRH, was only minimally effected by hperPRL at 3 and 7 days postcastration. LH responses were similar at all time points after GnRH in control and prolactin-treated animals, except for the peak LH responses, which were significantly smaller in the prolactin-treated animals. The effects of hyperPRL were examined further by exposing hemipituitaries in vitro from male rats to 6-min pulses of GnRH (5 ng/ml) every 30 min for 4 h. HyperPRL had no effect on basal LH release in vitro, on GnRH-stimulated LH release, or on pituitary LH concentrations in hemipituitaries from animals that were intact, 3 days postcastration, or 7 days postcastration. However, net GnRH-stimulated release of FSH was significantly higher by pituitaries from hyperprolactinemic, castrated males. To assess indirectly the effects of hyperPRL on GnRH release, males were subjected to electrical stimulation of the arcuate nucleus/median eminence (ARC/ME) 3 days postcastration. The presence of elevated levels of prolactin not only suppressed basal LH secretion but reduced the LH responses to electrical stimulation by 50% when compared to the LH responses in control castrated males. These results suggest that acute hyperPRL suppresses LH secretion but not FSH secretion. Although pituitary responsiveness is somewhat attenuated in hyperprolactinemic males, as assessed in vivo, it is normal when pituitaries are exposed to adequate amounts of GnRH in vitro. Thus, the effects of hyperPRL on pituitary responsiveness appear to be minimal, especially if the pituitary is exposed to an adequate GnRH stimulus. The suppression of basal LH secretion in vivo most likely reflects inadequate endogenous GnRH secretion. The greatly reduced LH responses after electrical stimulation in hyperprolactinemic males exposed to prolactin suggest further that hyperPRL suppresses GnRH secretion.     

Stimulation of the secretion of luteinizing hormone by ginsenoside-Rb1 in male rats

Ginsenoside-Rb1 is one of the pharmacologically active components of ginseng, the dry root of Panax ginseng C. A. Meyer (Araliaceae), a well-known traditional Chinese medicine. Ginseng enhanced mounting behaviour of male rats and increased sperm counts in rabbit testes. Some experimental results suggested no sex hormone-like function in ginseng but probably gonadotropin-like action. The present study was to explore the effect of ginsenoside-Rb1 on the secretion of luteinizing hormone (LH) both in vivo and in vitro. Male rats were orchidectomized (Orch) for 2 weeks or subjected to swim training for 1 week before catheterization via the right jugular vein. They were intravenously injected with ginsenoside-Rb1 (10 microg/kg) or saline at 15 min prior to a challenge of gonadotropin-releasing hormone (GnRH) or 10 min-swim. Blood samples were collected at several time intervals following intravenous injection of ginsenoside-Rb1. In the in vitro experiment, male rats were decapitated and their anterior pituitary gland (APs) were either bisceted or enzymatically dispersed. The hemi-APs were preincubated with Locke's medium at 37 degrees C for 90 min and then incubated with Locke's medium containing ginsenoside-Rb1 (10(-7) - 10(-4) M) for 30 min. The dispersed AP cells (1 x 1(5) cells per well) were primed with dihydrotestosterone (DHT, 10(-8) M) for 3 days, and then challenged with ginsenoside-Rbl (10(-6) and 10(-5) M, n = 8) for 3 h. The concentrations of LH or testosterone in samples were measured by radioimmunoassays. Administration of ginsenoside-Rb1 did not alter the levels of plasma LH in both intact and Orch rats but significantly increased plasma LH concentration at the termination of the 10 min swimming exercise. Administration of ginsenoside-Rb1 resulted in a lower testosterone response to GnRH challenge or swimming exercise as compared with saline-treated rats. Ginsenoside-Rb1 dose-dependently increased the release of LH from both hemi-AP tissues and the DHT-primed dispersed AP cells in vitro. These results suggest that ginsenoside-Rb1 increases LH secretion by acting directly on rat AP cells.     

Changes in the dynamics of luteinizing hormone-releasing hormone-stimulated secretion of luteinizing hormone during sexual maturation of female rats

Our aim was to identify age-related changes in the dynamics of luteinizing hormone (LH) release that may contribute to the decline in pituitary sensitivity to luteinizing hormone-releasing hormone (LHRH) during sexual maturation of female rats. We studied LHRH-stimulated LH secretion curves of superfused pituitaries from rats ranging in age from 10 days to the first estrous cycle. Pituitary fragments were exposed for 10 min to medium alone or to medium plus LHRH; incubation continued in medium alone for 130 min and effluent was collected for LH analysis. Secretion curves were compared on the basis of total secretion (area under the curve), maximal change in LH secretion rate, and rates of rise and decay of the curves. The data show that total LH secretion in response to LHRH is greatest in 15-, 20-day-old and first-proestrus animals. Also, the maximal change in LH secretion rate was greater, and the increase in LH secretion rate faster in younger animals than in 30-day-old animals. Analysis of secretory granules in LH-containing gonadotropes of 15- and 30-day-old animals revealed changes in he granule population with age. We conclude that younger animals respond faster with a greater LH secretion response to LHRH than do 30-day-old or first-estrus animals, and that these age-related changes in the dynamics of LH secretion may be due in part to maturation of the LH secretory granules.