Luteinizing Hormone-releasing Hormone Pulse Research Articles

Effects of transplants of fetal mediobasal hypothalamus on luteinizing hormone pulses impaired by hypothalamic deafferentation in adult ovariectomized rats.

Pulsatile luteinizing hormone (LH) secretion is impaired after posterior anterior-hypothalamic deafferentation (PAD), which separates the anterior part of the arcuate nucleus from the mediobasal hypothalamus (MBH). In the present study, we examined whether transplants of fetal brain tissue could prevent the effects of PAD. The brain tissue containing the MBH or the cerebral cortex taken from the fetal brain was transplanted into the third ventricle of ovariectomized rats. Four weeks after the brain transplantation, animals with or without the brain transplantation were subjected to PAD. One week after PAD, blood samples were collected every 6 min for 3 h through an indwelling atrial cannula. Rats bearing PAD without transplantation showed irregular pulsatile fluctuation of plasma LH, whereas LH pulses were maintained in rats bearing transplantation of the fetal MBH tissue. In rats which had been transplanted with the cerebral cortex, LH pulses were less apparent after PAD than in the MBH-transplanted or sham-deafferentated animals. No cell bodies of LH-releasing hormone (LHRH) neurons were found immunohistochemically in the MBH grafts. These results suggest that the graft containing the fetal MBH tissue maintains regular LH pulses after PAD and that the LHRH pulse generator may consist, at least in part, of a group of neurons in the MBH other than LHRH-producing neurons.

Estradiol and progesterone effects on relative luteinizing hormone and follicle stimulating hormone release induced from superfused anterior pituitary cell cultures by defined LHRH pulse regimens

These studies examined the capacity of estradiol and progesterone to modulate relative luteinizing hormone (LH) and follicle stimulating hormone (FSH) secretion from superfused anterior pituitary cells when stimulated with luteinizing hormone releasing hormone (LHRH) pulse regimens of specific amplitude, duration and frequency. There was particular interest in whether such steroid and LHRH treatments induced evidence of divergent LH or FSH secretion. Pituitaries were recovered from adult, 2 week ovariectomized rats and cultured for 48 h with collagen coated Cytodex microcarrier beads. Cultures were preincubated either with or without estradiol (1 or 10 nM) for 48 h and were subsequently incubated for 3, 6 or 12 h with 100 nM progesterone. All groups were then pulsed with 1 of 3 LHRH regimens; regimen 1 delivered 8 ng in a single 100 μl bolus once/h; regimen 2 divided the 8 ng dose of regimen 1 into 3 equal doses administered at 4 min intervals thereby maintaining the 8 ng mass of regimen 1 while extending the duration of exposure; regimen 3 was the same as regimen 2 except that the 3 equal doses were administered at a pulse frequency of 1 per 2 h rather than 1 per h thereby not only maintaining the duration of exposure as in regimen 2 but also reducing the pulse frequency. 1 nM estrogen alone for 48 h had no effect on LHRH stimulated LH release regardless of regimen; however, FSH was increased when hourly pulses of increased duration were applied (regimen 2). When estrogen was increased to 10 nM, regardless of regimen, LH was predominantly inhibited while FSH was unaffected. When 1 nM estrogen was followed by progesterone, both LH and FSH were elevated at 6 h progesterone in response to regimen 2; with 10 nM estrogen however, a divergent response was observed in that LH release was elevated at 6 h while FSH was elevated at 3 h in response to regimens 2 and 3. These results first of all confirm that progesterone in combination with estrogen is capable of exerting both inhibitory and stimulatory effects on gonadotropin secretion; secondly, these studies show that, as a direct pituitary effect, the LHRH regimen and the gonadal steroid milieu are capable of interacting to significantly influence the relative secretion of LH and FSH. The data therefore suggest that the divergent gonadotropin secretion seen in various physiological states in vivo is due likely in part to a combination of estrogen and progesterone priming in combination with the hypothalamic LHRH secretory pattern.

Pulsatile Luteinizing Hormone Responses to Intermittent iV-Methyl-D,L- Aspartate Administration in Hamsters Exposed to Long- and Short-Day Photoperiods*

Hamsters exposed to short-day photoperiods (6-h light, 18-h darkness) develop a reversible inhibition of the reproductive axis which includes decreased LH pulse frequency, decreased FSH and testosterone levels, testicular regression, and presumably, decreased luteinizing hormone-releasing hormone (LHRH) release. The decrease in LHRH release could reflect a decrease in the ability of LHRH neurons to release the decapeptide in response to intermittent neuronal excitation. To analyze this possibility, the LHRH secretagogue, N-methyl-D-aspartate (NMA) was used to estimate relative releasability of endogenous LHRH pulses in hamsters exposed to long- and short-day photoperiods. Long-day (LD) or short-day (SD) hamsters were fitted with indwelling atrial catheters 2 days before experimentation. During 5-h sampling sessions (1300-1800 h), blood was withdrawn at 10-min intervals. In initial experiments, a NMA dose-response analysis revealed that 10 and 20 mg/kg NMA but not 2.5 and 5 mg/kg NMA, produced significant, dose-related LH responses in LD hamsters. Treatments with LHRH antagonist 4 h prior to NMA treatment completely blocked LH responses. The 10 mg/kg NMA dose was then used to directly compare LHRH responsiveness in LD and SD hamsters. NMA injections were administered to groups of LD and SD hamsters after hours 1, 2, 3, and 4 of sampling. In LD hamsters, LH responses to the first NMA pulse (2.65 +/- 0.09) were followed by diminishing responses to the second, third, and fourth NMA pulses (1.22 +/- 0.50, 1.22 +/- 0.43, and 1.15 +/- 0.42). By contrast, initial LH responses in SD hamsters (1.61 +/- 0.31) were followed by even more robust LH responses to the second, third, and fourth NMA challenges (2.89 +/- 0.34, 3.08 +/- 0.59, 2.65 +/- 0.32). Although LH responses to the first NMA pulse were slightly less in SD vs. LD animals, responses to all subsequent injections were actually greater in SD hamsters vs. their LD counterparts. Injections of 20 ng/kg LHRH in SD hamsters produced LH responses similar to those evoked by NMA. The same LHRH dose also produced continued, robust LH responses in LD hamsters, indicating that decrements in response to NMA were not due to diminished pituitary responsiveness. These results suggest that 1) the stimulatory actions of NMA are mediated by LHRH, 2) exposure to short days does not decrease the responsiveness of LHRH neurons to NMA, and 3) successive NMA challenges produce diminishing LH responses in LD hamsters, but not in SD hamsters.(ABSTRACT TRUNCATED AT 400 WORDS)

Exogenous versus endogenous pulses of luteinizing hormone-releasing hormone and secretory patterns of gonadotropins

Does an exogenously administered regimen of luteinizing hormone-releasing hormone (LH-RH) pulses override endogenous LH-RH? Pulses of LH-RH were given intravenously during 1 week with intervals of 90 (n = 5) or 120 minutes (n = 5). Before, during, and after treatment serial plasma luteinizing hormone (LH) and follicle-stimulating hormone (FSH) patterns and pituitary responsiveness to LH-RH were estimated. Women with virtual absence of gonadal function (postmenopause, gonadal dysgenesis, and premature menopause). During treatment with the 90-minute interval, no LH pulses that were not related to injections of LH-RH were observed. Two spontaneous LH pulses were observed during treatment with the 120-minute interval. Immediately after treatment, a lowered incidence of spontaneous LH pulses was seen of 3 pulses/6 h if LH-RH had been given every 90 minutes and to 1.5 pulses/6 h after the 120-minute interval treatment. Gonadotropin responses to 100 μ g of LH-RH were attenuated during treatment but recovered within 48 hours after discontinuation of treatment. (1) Exogenously administered LH-RH can override endogenous LH-RH or its effects on the release of LH in women with hypergonadotropic hypogonadism; (2) during pulsatile LH-RH treatment desensitization of the pituitary occurs to some degree; and (3) immediately after cessation of treatment with pulsatile LH-RH, spontaneous LH pulses are present but with a significantly lower incidence.

The luteinizing hormone-releasing hormone pulse generator in men: Abnormalities and clinical management

Most men with hypogonadotropic eunuchoidism have absent luteinizing hormone and presumably absent luteinizing hormone-releasing hormone pulses. Pulsatile luteinizing hormone-releasing hormone therapy is effective in restoring normal gonadotropin secretion and testicular function and inducing fertility in men with hypogonadotropic eunuchoidism. Furthermore, pulsatile (versus continuous) luteinizing hormone-releasing stimulation of the pituitary gland is an absolute requirement for normal gonadotropin secretion. Men with idiopathic oligoazoospermia and selective elevation of follicle-stimulating hormone levels have slow luteinizing hormone and presumably luteinizing hormone-releasing pulse frequency. In these men, pulsatile luteinizing hormone-releasing treatment is effective in decreasing serum follicle-stimulating hormone levels, but it is unclear whether spermatogenesis and fertility are improved. (AM J OBSTET GYNECOL 1990;163:1743–52.)

The induction of divergent gonadotropin secretion in vitro by variations in luteinizing hormone releasing hormone pulse regimen

The effects of luteinizing hormone releasing hormone (LHRH) pulse amplitude, duration, and frequency on divergent gonadotropin secretion were examined using superfused anterior pituitary cells from selected stages of the rat estrous cycle. Cells were stimulated with one of five LHRH regimens. With low-amplitude LHRH pulses (regimen 1) in the presence of potentially estrogenic phenol red, LH response in pituitary cells from proestrus 1900, estrus 0800, and diestrus 1 0800 were all significantly larger(P < 0.05) than the other stages tested. In the absence of phenol red, responsiveness at proestrus 1900 was significantly larger than proestrus 0800, proestrus 1500, and estrus 0800 (P < 0.01, 0.05, and 0.05, respectively); other cycle stages tested were smaller. No significant differences were observed between cycle stages for follicle-stimulating hormone (FSH) secretion in the presence or absence of phenol red. Because pituitary cells at proestrus 1900 were the most responsive to low-amplitude 4 ng LHRH pulses, they were also used to study the effects of LHRH pulses of increased amplitude or duration and decreased frequency. Increasing the amplitude (regimen 2) or the duration (regimens 3 to 5) increased FSH secretion; this effect was greatest with regimens 3 and 5. When regimens 3 and 5 were studied in pituitary cells obtained at proestrus 1500, FSH was significantly increased by both regimens, but most by regimen 5; furthermore, LH release was significantly reduced. When regimens 3 and 5 were studied in pituitary cells obtained at estrus 0800, FSH release was elevated most significantly by regimen 5. Thus, variations in LHRH pulse regimen were found to he capable of inducing significant divergence in FSH release from superfused anterior pituitary cells derived from specific stages of the estrous cycle.

Seasonal changes in the in-vivo activity of the luteinizing hormone-releasing hormone (LHRH) neural apparatus of male rabbits monitored with push-pull cannulae

Intact sexually mature New Zealand White male rabbits, raised under natural lighting and temperature conditions, were isolated and housed in air-conditioned quarters, in a 12-h light:12-h dark cycle. Push-pull cannulae were implanted towards the tuberal region of the hypothalamus, and animals were perfused with modified Krebs'-Ringer phosphate medium for an average period of 4 h. Most rabbits were repetitively perfused over an average period of 3.7 months. Perfusions were grouped into seasonal periods of about 40 or 80 days through the entire 1-year cycle: (A) 23 November to 31 December (winter solstice period: N = 6), (B) 1 January to 23 March (winter: N = 10), (C) 24 March to 13 June (spring: N = 9), (D) 14 June to 23 July (summer solstice period: N = 23), (E) 24 July to 13 October (late summer-early fall: N = 7) and (F) 14 October to 22 November (fall: N = 4). Maximal and minimal values of mean release, mean amplitude and mean frequency from every animal in each block were obtained. In the summer solstice group (D), maximal mean (+/- s.e.) LHRH release levels were significantly greater (14.42 +/- 6.62 pg/10 min) than for all groups (A, 0.76 +/- 0.27; B, 1.59 +/- 0.39; C, 1.34 +/- 0.22; E, 1.33 +/- 0.33; and F, 1.18 +/- 0.11) while during the winter solstice period (A), minimal mean LHRH release levels (0.48 +/- 0.04 pg/10 min) were significantly lower than in all other groups (B, 1.43 +/- 0.41; C, 0.96 +/- 0.11; D, 7.25 +/- 4.12; E, 1.18 +/- 0.37; and F, 1.18 +/- 0.11). Maximal values were highest during the summer solstice period while the minimal values were lowest during the winter solstice period. The amplitude and frequency of the LHRH pulses showed changes similar to those observed with the mean LHRH release. For 1 year, an estimated left testis weight was measured in 4-7 rabbits every 2-5 weeks. The percentage estimated testis weight peaked in early August and reached minimal levels during the winter months. These data demonstrate that the rabbit LHRH neural apparatus is very sensitive to seasonal influences although animals remained in a fixed photoperiod during the entire duration of the experiment.

Interactions between the Noradrenergic and Opioid Peptidergic Systems in Controlling the Electrical Activity of Luteinizing Hormone-Releasing Hormone Pulse Generator in Ovariectomized Rats

The effects of noradrenergic and opioid peptidergic receptor blockade, either alone or in combination, on the electrical activity of luteinizing hormone-releasing hormone (LHRH) pulse generator were studied in ovariectomized rats fitted with chronically implanted electrode arrays in the medial basal hypothalamus. Both alpha- and beta-adrenergic receptor antagonists, i.e. phenoxybenzamine (5 mg/kg i.v.) and propranolol (5 mg/kg i.v.), respectively, significantly increased the intervals between characteristic increases (volleys) in hypothalamic multiunit activity (MUA), which were associated with the initiation of LH pulses. In contrast to this, an opioid receptor antagonist naloxone (2 mg/kg i.v.) significantly decreased the intervals between the MUA volleys. Naloxone given after the injection of propranolol induced MUA volleys with a latency of a few minutes. However, when given after the injection of phenoxybenzamine, naloxone failed to induce such immediate MUA volleys. These changes in the intervals between the MUA volleys were faithfully reflected by the pulsatile LH secretion. These results suggest that norepinephrine facilitates LHRH pulse generator activity through both alpha- and beta-adrenergic-receptors, and that the action of opioid peptides on it requires an alpha-adrenergic receptor-mediated mechanism.

Effect of Mating Behavior on Luteinizing Hormone-Releasing Hormone Release in Female Rabbits as Monitored with Push-Pull Cannulae

To examine the effect of mating behavior on luteinizing hormone-releasing hormone (LHRH) release, intact New Zealand female rabbits were implanted with push-pull cannulae (PPC) aimed at the tuberal region of the hypothalamus and perfused with modified Krebs-Ringer phosphate medium at 11-13 microliters/min. In the mating experiments, does (n = 10) were initially perfused for a control period of 60-170 min followed by a mating period (100-160 min) which included the introduction of the male rabbit for an average time period of 30 min. Two groups of LHRH release patterns were observed: positive and negative responders. In the positive LHRH responders (n = 5), a clear rapid increase in LHRH release following mounting by the male occurred with a significant increase in the mean LHRH release (1.83 +/- 0.33 to 3.27 +/- 0.80 pg/10 min, p less than 0.040), in the mean LHRH amplitude (1.97 +/- 0.46 to 4.33 +/- 1.29 pg, p less than 0.022) and in the amplitude of the largest LHRH pulse (2.13 +/- 0.43 to 7.58 +/- 3.65 pg, p less than 0.022). In the negative LHRH responders (n = 5), no changes in LHRH release were detected although all rabbits ovulated, with some becoming pregnant. It appears from histological analysis that the difference between these two patterns of responses following mating are due to different cannula placements. In the positive responders, the tip of the PPC was localized in the tuberal region whereas in the negative responders, the placements were more dorsal and, in some cases, anterior.(ABSTRACT TRUNCATED AT 250 WORDS)

Naloxone Increases the Frequency of the Electrical Activity of Luteinizing Hormone-Releasing Hormone Pulse Generator in Long-Term Ovariectomized Rats

Characteristic increases in neuronal activity accompanied by the initiation of each luteinizing hormone (LH) pulse have been successfully recorded in freely moving ovariectomized rats by means of multiunit activity (MUA) recording techniques. In the present study, the effect of the opioid receptor antagonist naloxone on this neuronal activity was examined in rats ovariectomized for 6-10 weeks. Electrodes were chronically implanted into the medial basal hypothalamus (n = 78) or the medial preoptic area (n = 29). During the MUA recording, blood samples were taken through an indwelling atrial cannula at 3- or 6-min intervals to determine the serum LH concentration. Naloxone was administered intravenously for 1 h by either intermittent injection every 6 min (0.05 mg/kg, 10 times) or continuous infusion (0.1, 0.2 and 0.5 mg/kg/h). Explosive rises in the MUA (volleys) associated with the initiation of LH pulse were recorded in 7 animals in which the tips of electrodes were located in the arcuate nucleus-median eminence region. In 5 animals studied, the mean (+/- SE) duration and interval of the MUA volleys were 2.1 +/- 0.1 and 22.1 +/- 0.9 min, respectively. Naloxone given every 6 min significantly increased the duration to 2.6 +/- 0.1 min, and decreased the interval to 9.5 +/- 0.9 min. This naloxone treatment disturbed the clear pulsatility of LH secretion observed in the pretreatment control period. Analysis of the effects of naloxone infused at 3 different doses revealed that the facilitatory effects of naloxone on the duration and frequency of the MUA volleys were dose-dependent.(ABSTRACT TRUNCATED AT 250 WORDS)

The luteinizing hormone-releasing hormone pulse generator in men: Abnormalities and clinical management

Most men with hypogonadotropic eunuchoidism have absent luteinizing hormone and presumably absent luteinizing hormone-releasing hormone pulses. Pulsatile luteinizing hormone-releasing hormone therapy is effective in restoring normal gonadotropin secretion and testicular function and inducing fertility in men with hypogonadotropic eunuchoidism. Furthermore, pulsatile (versus continuous) luteinizing hormone-releasing stimulation of the pituitary gland is an absolute requirement for normal gonadotropin secretion. Men with idiopathic oligoazoospermia and selective elevation of follicle-stimulating hormone levels have slow luteinizing hormone and presumably luteinizing hormone-releasing pulse frequency. In these men, pulsatile luteinizing hormone--releasing treatment is effective in decreasing serum follicle-stimulating hormone levels, but it is unclear whether spermatogenesis and fertility are improved.

The frequency of pulsatile luteinizing hormone-releasing hormone treatment and luteinizing hormone and follicle-stimulating hormone secretion in women with amenorrhea of suprapituitary origin

The influence of luteinizing hormone-releasing hormone (LH-RH) pulse frequency on luteinizing hormone (LH) and follicle-stimulating hormone (FSH) was studied in hypogonadotropic hypogonadal women. They received three regimens of 5 days of pulsatile LH-RH (5 micrograms/pulse) given at 30-, 90-, or 180-minute intervals, with at least 6 weeks between treatments. On day 1, LH and FSH increased in proportion to the LH-RH pulse frequency. After 5 days of treatment with the 30- and 90-minute intervals, LH was still elevated, but FSH had returned to pretreatment levels together with a decline of the FSH response. The LH response only declined during treatment with the 30-minute pulse interval. During each treatment, estradiol (E2) increased. Explanations for dissociation between LH and FSH secretion during treatment with higher LH-RH pulse frequencies could be: (1) desensitization of FSH rather than LH secretion on LH-RH; (2) a differential effect of E2 on LH and FSH; (3) nonsteroidal ovarian factors selectively regulating LH and/or FSH release.

Serotonin stimulation of the period of in vitro LHRH release is estradiol dependent

The neurotransmitter serotonin (5-HT) has well known effects on the release of luteinizing hormone (LH) which may be estrogen and photoperiod dependent. An in vitro superfusion system was used to assess the potential role of 5-HT and photoperiod in modulating the release of luteinizing hormone releasing hormone (LHRH) from the mediobasal-suprachiasmatic-preoptic area (MBH-POA-SCN) in the ovariectomized (OVX) and OVX estradiol (E 2) implanted rat model of LH release. Regularly cycling female Holtzman strain rats, maintained on a photoperiod of 0500–1900 or 0100–1500 hr light were OVX, and silastic capsules containing estradiol-17 beta (150 μg/ml) or sesame oil implanted SC. Two days later the rats were killed at 1000 hr and the MBH-POA-SCN isolated. Tissue was superfused in vitro with Krebs-Ringer-Phosphate buffer (KRP) for 2 hours followed by 3 hours of KRP with intermittent superfusion of 5-HT at increasing concentrations: 1 × 10 −10 M (1st hour), 1 × 10 −8 (2nd hour), 1 x 10 −6 M (3rd hour). Superfusate was collected every 10 min for the entire 5 hour incubation period and LHRH measured by RIA with the following results. In the OVX rat 5-HT significantly ( p 0 ̌ .05 ) increased the period of LHRH pulses and decreased the release of LHRH compared to control superfusions, while in the OVX E 2 rat 5-HT significantly ( p 0 ̌ .05 ) decreased the period of LHRH release and increased the release of LHRH. In another series of experiments the tissue was perfused for 2 hours with KRP, 2 hours with 1 x 10 −8 M 5-HT, and 2 hours with KRP. 5-HT again significantly increased ( p 0 ̌ .05 ) the period of LHRH pulses from tissue derived from the OVX rat and decreased the period ( p 0 ̌ .05 ) of LHRH pulses in tissue from the OVX E 2-treated rat. However, in the OVX-treated rat LHRH release was also increased ( p 0 ̌ .05 ) during the first hour of 5-HT superfusion. Photoperiod had no significant effect on the release of LHRH from OVX- and OVX E 2-treated rats. However, the effect of 5-HT in modulating the frequency of LHRH release may be related to the presence of E 2 in the rat and could serve as one mechanism in vivo to coordinate the release of LHRH with endocrine cues.

Steroid receptors and regulation of LH and secretogranin II (GP87) secretion in pituitary cell aggregates

The gonadotrope polypeptide (GP87), a secretogranin II form, has previously been found to be co-released with luteinizing hormone (LH) by rat gonadotrope cells under luteinizing hormone-releasing hormone (LHRH) stimulation. Nothing is hitherto known concerning its function and regulation. In the present paper, the modulatory effects of steroids on GP87 release were investigated using cultures of rat pituitary cell aggregates. As steroid effects are dependent on the presence of receptors, we firstly demonstrated that the steroid receptors are maintained with their typical characteristics in this culture system. In steroid-free medium, the estrogen receptor concentration increased significantly (P < 0.01) within 7 days in culture (646 +/- 122 fmol/mg DNA, n = 3) when compared to the level in freshly dispersed cells (355 +/- 59 fmol/mg DNA, n = 4) suggesting that cells can synthesize the estrogen receptor and that this cell culture system is suitable for studying regulatory effects of steroids. Pituitary cell aggregates from 14 day-old female rats were maintained for 24 h in the absence or presence of 10 nM estradiol (E(2)) or 50 nM 5?-dihydrotestosterone (DHT) and incubated for 2.5 h with [(35)S] methionine. Perifusion was initiated and 4 min pulses of 20 nM LHRH occurred every 1 h. The effects of steroids were investigated on LH, total and labeled GP87. The results indicated that: (1) DHT pretreatment reduced the amplitude of LHRH-stimulated GP87 and LH release responses; (2) by contrast, E(2) pretreatment enhanced the effects of LHRH on total or labeled GP87 release whereas the LH release was not significantly modified; and (3) when E(2) treatment started only 60 min before the first LHRH pulse, no differences between responses of E(2)-treated and control aggregates were observed. The present data show for the first time that sex steroids can modulate the GP87 release through a direct effect on the pituitary. E(2) could either stimulate the GP87 synthesis or increase its intracellular trafficking. That LH and GP87 do not strictly exhibit the same release response further suggests either some heterogeneity of GP87 localization in gonadotrope sub-types or within secretory granules.

Inhibitory effect of central LHRH on LH secretion in the ovariectomized ewe.

The role of central luteinizing hormone releasing hormone (LHRH) in the control of pulsatile LHRH and luteinizing hormone (LH) secretion was investigated in ovariectomized adult ewes. Injection of LHRH (2.1-21 pmol) into the third cerebral ventricle caused a delayed but sustained inhibition of LH secretion. Pulse frequency, pulse amplitude and mean LH levels were reduced significantly when compared with the responses to the control injection of saline (50 microliters). The inhibitory effect of centrally administered LHRH was not accompanied by a reduction in the pituitary responsiveness to intravenous LHRH. In contrast to the effect on LH, plasma levels of follicle-stimulating hormone (FSH) and prolactin were unaffected by central LHRH. The inhibitory action of LHRH was antagonized by prior injection of an LHRH antagonist ([N-Ac-D-Nal(2)1, D-p-Cl-Phe2, D-Trp3, D-hArg (Et2)6, D-Ala10] LHRH, 69 pmol) into the third ventricle. Central injection of the LHRH antagonist alone (at the same concentration) did not influence any characteristic of pulsatile LH secretion. In conclusion, these data indicate that exogenous administration of LHRH into the brain exerts a dose-related and receptor-mediated inhibition of LHRH pulse generator activity. However, the physiological significance of endogenous LHRH in the regulation of the LHRH pulse generator remains unresolved.

Alterations in Luteinizing Hormone-Releasing Hormone Release from the Mediobasal Hypothalamus of Ovariectomized, Steroid-Primed Middle-Aged Rats as Measured by Push-Pull Perfusion

The spontaneous luteinizing hormone (LH) surge on proestrus as well as the steroid-induced LH surge and hypersecretion of LH following ovariectomy are attenuated in middle-aged female rats when compared to their young counterparts. It is generally assumed that the lower titers of serum LH measured in aging animals result, in part, from age-related alterations in luteinizing hormone-releasing hormone (LHRH) neurosecretion, yet no direct measurements of LHRH release from the hypothalamus of aging females are currently available. The present study utilized the push-pull perfusion technique to characterize and compare in vivo LHRH output from the mediobasal hypothalamus of ovariectomized middle-aged and young females during a steroid-induced LH surge. Twelve-minute perfusates were collected for a period of 6 h from middle-aged and young animals outfitted with push-pull cannulae resting in the mediobasal hypothalamus in close proximity to the median eminence. The overall pattern of LHRH release differed in the two age groups. Mean levels of LHRH measured in brain perfusates from middle-aged females were significantly lower than mean levels measured in young animals. Moreover, mean levels of LHRH detected in perfusates during the 1- and 2-hour intervals prior to and the 1-hour interval after the time of maximal LHRH output were lower in middle-aged animals, and a larger percentage of samples from middle-aged females contained levels of LHRH below the detectability of the assay. The mean amplitude of identified LHRH pulses and maximal LHRH output, however, did not differ in the two age groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Spontaneous changes in LHRH release during the rat estrous cycle, as measured with repetitive push-pull perfusions of the pituitary gland in the same female rats.

In the present study, we examined the in vivo release profiles of luteinizing hormone-releasing hormone (LHRH) during the rat estrous cycle by utilizing repetitive push-pull perfusions of the anterior pituitary in the same freely behaving animal. Throughout the estrous cycle, LHRH levels were higher than those reported from previous work using a push-pull cannula placed in the mediobasal hypothalamus. A significant increase in LHRH input to the anterior pituitary was obtained in proestrus (Pro), though the magnitude of increment was different among animals. This LHRH increment in Pro was attributable to a significant increase in the amplitude of LHRH pulses. Interestingly, no changes were observed in the frequency of LHRH pulses (about 1 pulse/50 min) throughout the estrous cycle. The data demonstrate that the anterior pituitary of female rats receives an increased amount of LHRH during Pro due to changes in the amplitude of the LHRH signal without changes in the frequency.

Luteinizing hormone releasing hormone (LHRH) neuroterminals mapped using the push-pull perfusion method in the rhesus monkey

In vivo luteinizing hormone releasing hormone (LHRH) release was measured in conscious, ovariectomized rhesus monkeys using a push-pull cannula inserted into the stalk-median eminence, and the relationship between sampling location and LHRH release was examined. Within an individual animal in which multiple experiments were conducted with different cannula placements, LHRH pulse frequency was consistent. In contrast, LHRH pulse amplitude and mean LHRH release varied with cannula tip location in a pattern which reflected the anatomical distribution of LHRH-immunoreactive fibers described for the rhesus monkey. These results suggest that our push-pull perfusion method is reliable for the in vivo measurement of LHRH and perhaps other neuropeptides and/or neurotransmitters, as well.

Modulation of luteinizing hormone pulse amplitude by the frequency of luteinizing hormone-releasing hormone stimulation and by testosterone in castrated, hypothalamic-lesioned male rats.

The frequency of spontaneous luteinizing hormone (LH) pulses is thought to be a direct result of the frequency of luteinizing hormone-releasing hormone (LHRH) pulses from the hypothalamus. By contrast, the amplitude of spontaneous LH pulses may be controlled by several factors other than the amplitude of LHRH pulses. We tested two hypotheses: 1) that LH pulse amplitude is determined in part by the frequency of LHRH pulses of constant magnitude, and 2) that testosterone (T) exerts a direct feedback effect on the pituitary gland to regulate LH pulse amplitude. Gonadal feedback was eliminated by castrating adult male rats (n = 20). Endogenous LHRH secretion was eliminated by lesioning the medial basal hypothalamus. Serum LH levels (0.19 +/- 0.04 ng/ml RP-2, mean +/- SEM) and T levels (0.15 +/- 0.02 ng/ml), measured several weeks after hypothalamic lesioning, confirmed the hypogonadotropic hypogonadal state of the animals. During a 8-h period, unanesthetized, unrestrained animals were injected with 40-ng pulses of LHRH via catheters into the jugular vein, and blood samples for LH measurement were drawn at 10-min intervals. The LHRH pulse interval was 20 min during the first 4 h in all animals. The pulse interval was doubled to 40 min in half of the animals (n = 10) during the next 4 hours; in the other 10 animals, the pulse interval was maintained constant at 20 min throughout the study. Within both of these groups, one-half of the animals (n = 5) were infused with T to achieve a physiological level of T in serum (2.46 +/- 0.36 ng/ml at 4 h), while the other half received vehicle.(ABSTRACT TRUNCATED AT 250 WORDS)

The effect of estrogen on episodic secretory patterns of luteinizing hormone-releasing hormone (LHRH) and luteinizing hormone (LH) in hypergonadotropic hypogonadism

The secretory dynamics of plasma luteinizing hormone-releasing hormone (LHRH) and serum luteinizing hormone (LH) were studied in three hypogonadal women before and after chronic administration of mestranol. Blood samples were obtained through an indwelling iv line every 15 min over 3 hours, and plasma levels of LHRH and LH were measured by radioimmunoassay. LHRH and LH pulses were defined as rising from nadir to peak that exceed 2 times the intraassay coefficient of variation. All patients showed pulsatile LHRH and LH release before mestranol administration. The mean LH levels (89 +/- 20 mIU/ml) and pulse amplitude (33 +/- 14 mIU/ml) were significantly reduced after mestranol administration. On the other hand, the mean LHRH levels (1.87 +/- 0.49 pg/ml) and pulse amplitude (0.92 +/- 0.41 pg/ml) did not change significantly after mestranol administration. Pulse frequency (2 approximately 3 times/3 hrs) of LHRH and LH did not change after mestranol administration. These data show that the chronic administration of estrogen to such patients cause a decrease in mean LH levels and amplitude of LH pulse without a decrease of pulsatile LHRH secretions. These results suggest that the chronic negative feedback action of estrogen on episodic LH release in women may be at the level of the pituitary gland and estrogen may change the pituitary sensitivity to LHRH.