Pituitary Luteinizing Hormone Content Research Articles

Pituitary and ovarian responses to luteinizing-hormone-releasing hormone during pregnancy and after parturition in brown hares (Lepus europaeus)

Female hares were given an i.v. injection of 5 micrograms luteinizing-hormone-releasing hormone (LHRH) between Days 7 and 19 (n = 21), 20 and 33 (n = 17) and 34 and 41 (n = 17) of pregnancy, and in the 3 days after parturition (n = 16). Whatever the stage of pregnancy, the LHRH injection induced a release of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) and an acute secretion of progesterone; these hormonal responses increased significantly during pregnancy, to reach values similar to those observed in nonpregnant, nonpseudopregnant females during the breeding season in the 3 days after parturition. However, the release of LH remained monophasic in pregnant and post-partum females, in contrast to the unmated females during the reproductive season, in which there was a biphasic profile. The proportion of ovulating females after LHRH treatment was approximately 60% at the beginning and end of pregnancy; and, after parturition, fell to 23% between Days 20 and 33. After Day 33, the pituitary response to LHRH was significantly higher in ovulating than in nonovulating females. At the beginning of pregnancy, 67% of females aborted after LHRH injection; after Day 20, the incidence of abortion decreased significantly and was 0% from Day 34. The amplitude and duration of progesterone secretion by the new corpora lutea resulting from ovulation after LHRH injection were similar to those of corpora lutea induced in nonpregnant females during the breeding season.(ABSTRACT TRUNCATED AT 250 WORDS)

A Role forN-Methyl-D-Aspartate Receptors in the Control of Seasonal Breeding*

The neuroendocrine basis for short-day induced testicular regression was studied in Syrian hamsters of the LSH/Ss Lak strain. Adult animals were maintained either under long or short days (14:10D or 6L:18D, respectively) and given single, daily ip injections of N-methyl-D-aspartate (NMDA) (25 mg/kg BW); control animals received injections of saline. As expected, the testes of the short-day controls had completely regressed to a prepubertal condition within 6 weeks, a change that was associated with significantly reduced mean plasma gonadotropin levels. In contrast, the NMDA-treated hamsters from both the long-day and short-day groups, as well as the long-day controls, all maintained large testes and elevated plasma gonadotropin levels, although plasma luteinizing hormone (LH) was partially suppressed in the short-day group. In a second experiment which lasted 2 weeks, short-day hamsters with completely regressed testes were either transferred to long days, maintained further on short days, or maintained on short days and given a daily ip injection of NMDA (25 mg/kg BW). The short-term exposure to long days caused an expected increase in plasma and pituitary concentrations of follicle-stimulating hormone, pituitary LH, and testicular weight. Similar, but even more marked, changes were observed in the short-day hamsters that were treated with NMDA, including significant increases in plasma LH and serum testosterone concentrations. Moreover, histological examination revealed that the recrudescing testes from this latter group already contained mature spermatocytes and in some individuals even spermatozoa. These results demonstrate that NMDA receptors may play a pivotal role in both the termination and onset of the breeding season in photoperiodic species.

2,3,7,8-Tetrachlorodibenzo- p-dioxin increases the potency of androgens and estrogens as feedback inhibitors of luteinizing hormone secretion in male rats

2,3,7,8-Tetrachlorodibenzo- p-dioxin (TCDD) decreases plasma androgen concentrations in male rats, without increasing plasma luteinizing hormone (LH) concentrations. If plasma LH concentrations had increased appropriately, plasma androgen concentrations in these animals would have returned to normal. The mechanism by which TCDD prevents the compensatory increase in plasma LH concentrations was therefore investigated. TCDD was found to have no effect on the plasma disappearance of iv administered LH. Therefore, the failure of plasma LH concentrations to rise was not due to increased clearance of LH from the circulation, but rather to an effect of TCDD on LH synthesis and/or secretion by the pituitary. In the absence of gonadal steroids (i.e., in castrated rats) TCDD did not prevent the compensatory increase in plasma LH concentrations from occurring. This was shown by 20-fold increases in plasma LH concentrations in both control and TCDD-treated rats 1 week after castration. Thus, (1) the presence of gonadal steroids is required for TCDD to prevent the compensatory increase in plasma LH concentrations, and (2) TCDD does not impair LH secretion by acting, itself, as an androgen or estrogen. TCDD treatment also did not affect pituitary LH content in castrated, testosterone-implanted rats. The above findings demonstrate that TCDD does not decrease the maximum rate at which the pituitary can synthesize and secrete LH. Rather, TCDD alters the feedback regulation of LH secretion when gonadal steroids are present. To determine if TCDD affects the potency of testosterone and its metabolites 5α-dihydrotestosterone and 17β-estradiol as feedback inhibitors of LH secretion, rats were dosed with TCDD, castrated, and implanted with sustained-release capsules containing graded amounts of each steroid. Seven days later, the potencies of all three hormones as feedback inhibitors of LH secretion were increased by TCDD, with little effect on their plasma concentrations. The TCDD dose dependence for the increased effectiveness of testosterone as a feedback inhibitor of LH secretion (ED50 10 μg/kg) was similar to that reported for the imbalance between plasma LH and androgen concentrations (ED50 15 μg/kg). Also, time courses for both responses were similar; each was detected within 1 day of TCDD dosing and each was fully developed after 7 days. We conclude that the mechanism by which TCDD prevents the compensatory increase in plasma LH concentrations in male rats is by increasing the potencies of androgens (and estrogens) as feedback inhibitors of LH secretion.

Gonadotropic dysfunction produced by Trypanosoma brucei brucei in the rat

Hormonal disorders have been frequently observed in humans and animals infected with tsetse-transmitted (African) trypanosomes. We studied the pituitary gonadal axis (plasma concentrations of testosterone, luteinizing hormone (LH), follicle stimulating hormone (FSH) and the pituitary gonadotropin (LH, FSH) concentrations) in rats as an experimental model infected with an acute stock of Trypanosoma brucei brucei (AnTat 1.1A). The same investigations in vivo were carried out with rats inoculated by trypanosomal preparations: surface coat components slowly released at pH 5.5 and the parasitic cellular pellet. The releasing procedure as firstly described by Baltz et al. (1976) was performed in the presence or the absence of protease inhibitors. We noted a testicular hypogonadism produced by the acute infection with the decrease of the testosterone level and an increase of the pituitary LH concentration, although the other circulating FSH and LH hormone levels were stable. The injection of the trypanosomal pellet, obtained in the presence of antiproteases, generated a similar clinical hormonal picture: decrease of testosterone level; increase in pituitary LH, FSH content; absence of significant variation of circulating FSH and LH rates. When the trypanosomal pellet was prepared in absence of antiproteases the circulating gonadostimuline levels were significantly decreased. In the same conditions (absence of antiproteases) the trypanosomal supernatant pH 5.5 induced the decrease of the testosterone and plasma LH levels. These results suggested that component(s) from trypanosomes generated hormonal perturbations.

Neuroendocrine Function in Transgenic Male Mice with Human Growth Hormone Expression

The neuroendocrine effects of human growth hormone (hGH) secretion were studied in adult male mice into which an hGH gene fused with mouse metallothionein 1 (mMT-1) promoter had been introduced. Intact transgenic mice had significantly greater plasma luteinizing hormone (LH) levels than did normal littermate controls. Castration increased LH levels in normal mice but was without effect on plasma LH levels in the transgenic mice. In vitro LH secretion and pituitary LH content were higher in the intact transgenic mice than in intact controls, while there was no significant difference in pituitary LH levels and in vitro LH secretion between the 2 groups of castrate animals. Intact transgenic mice exhibited a greater median eminence (ME) norepinephrine (NE) turnover than control animals, but ME NE turnover did not increase after castration in the transgenic animals as was the case in control mice. Castrate mice expressing the hGH gene had plasma levels of prolactin (PRL) similar to those seen in castrate controls, which was unexpected based on a previous study showing greatly attenuated PRL levels in intact hGH mice when compared to intact controls from the same line. Dopamine (DA) turnover in the ME was not significantly affected by the presence of the hGH gene, suggesting that the difference in plasma PRL levels between normal and transgenic mice is mediated through changes in PRL-regulating factors other than DA. In conclusion, the expression of the mMT-1/hGH hybrid gene in male mice leads to major alterations in LH secretion and lesser changes in PRL secretion.(ABSTRACT TRUNCATED AT 250 WORDS)

Reduction in Testicular Function in Rats

Chronic administration of dexamethasone in drinking water to maternal rats from days 15 to 21 of gestation (1) reduced plasma testosterone concentrations in male fetuses between days 19 and 21 but not earlier on day 18 and abolished the prenatal peak of plasma testosterone which normally occurs on day 19 of gestation, and (2) suppressed the postnatal surge of plasma testosterone in male newborns 1.5 and 2 h after delivery at term by cesarean section. The administration of dexamethasone to male fetuses at birth induced 1 h later a slight but not significant increase in hypothalamic gonadotropin-releasing hormone (GnRH) and pituitary luteinizing hormone (LH) contents, reduced drastically plasma LH levels and completely prevented the postnatal surge of plasma testosterone which occurred normally in littermate controls. A rise in pituitary LH content, and a sharp reduction in plasma LH and testosterone concentrations were noted in 19-day-old male fetuses whose mothers were acutely treated with dexamethasone on day 18 of gestation. Similar evolutions for LH were observed in littermate females. These results suggest that the inhibitory effects of exogenous glucocorticoids on testosterone secretion could be mediated in both fetuses and newborns at least partially through suppression of the hypothalamic and pituitary secretion of GnRH and LH, respectively, and provide insight how stress or hormone imbalance may affect the development of this neuroendocrine system.

Reduction in Testicular Function in Rats

A gonadotropin-releasing hormone (GnRH) antagonist, when injected 24 h before sacrifice to rat fetuses, did not modify plasma testosterone concentrations in males on day 18 of gestation but it did on days 19, 20 and 21. This GnRH antagonist reduced plasma luteinizing hormone (LH) levels and increased pituitary LH content in both male and female 19-day-old fetuses from mothers adrenalectomized on day 14 of gestation. An inverse relationship between plasma testosterone and LH levels was noted in males and females, on days 19 and 21. These data suggest that the hypothalamic control of gonadotropic function is operating by day 19 of fetal life and that a negative feedback of testosterone on LH and probably GnRH release is also operating in rat fetuses on days 19 and 21 of gestation.

Pubertal changes in the secretion of gonadotropic hormones, testicular gonadotropic receptors and testicular function in the ram

Developmental changes in pituitary content and secretory patterns of luteinizing hormone (LH), follicle-stimulating hormone (FSH) and prolactin (PRL), testicular size and steroidogenic function, testicular LH- and FSH-binding activity, and growth of the accessory sex organs were examined for 24 Dorset × Leicester × Suffolk rams (born in March) every 30 days from 30 to 150 days of age, and again at 200 days. Pituitary LH and FSH contents increased between 30 and 60 days of age and remained constant until 150 days, when contents were somewhat greater than on either 120 or 200 days. LH-pulse amplitude and frequency, and mean FSH concentration, were highest at 60 and(or) 90 days of age. Testicular growth increased dramatically between 90 and 150 days of age in association with increases in the number of LH- (100-fold) and FSH- (33-fold) binding sites in the testis and a small increase in blood testosterone concentration (1 ng/ml). During the same period, pituitary content and blood concentration of PRL increased to maximal values, epididymal, vesicular gland and bulbourethral gland weights increased 6-fold, and body weight doubled. Between 150 and 200 days of age, testosterone concentration increased considerably (8 ng/ml), as did LH-pulse frequency and the amount of LH- and FSH- binding in the testis; the reproductive organs continued to grow at a rate faster than that of the body as a whole. Testicular development of ram lambs was accompanied by increases in the secretion of all three pituitary hormones with gonadotropic properties, and in the number of LH and FSH receptors.

Eftects of Gonadectomy and Steroids on Pituitary Gonadotropin Secretion in a Frog, Rana Pipiens1

Secretory dynamics of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) were measured at various times following gonadectomy in adult male grass frogs, Rana pipiens. Plasma levels and in vitro initial secretory rates of both LH and FSH increased significantly within 1 wk and remained elevated for 3-4 wk of castration. Pituitary FSH and LH content were unchanged. However, dissociation between the two gonadotropins (Gth) occurred thereafter: Secretion of FSH remained elevated for 70 days, but those of LH declined to control levels after 30 days. In vitro secretion of Gth from gonadectomized (gonadx) frogs declined progressively over time reaching control levels after 24 h incubation. The results indicate that elevated pituitary secretion contributes to the observed circulating LH and FSH levels in gonadx frogs, and that FSH and LH may be controlled independently. Replacement therapy with 17 beta-estradiol (E2) suppressed post-gonadectomy increases in plasma Gth and in vitro responses to GnRH, whereas 5 alpha-dihydrotestosterone (DHT) had little effect in vivo and augmented GnRH responses in long-term castrates. In vitro, E2 also inhibited, while 48 h of DHT treatment had no effect on GnRH responsiveness of pituitaries from gonadx frogs. The actions of these steroids were opposite to those typically observed in mammals (and birds), and support the hypothesis that E2 may contribute to seasonal testicular regression in ranid frogs.

Testosterone and Dihydrotestosterone, but Not Estradiol, Selectively Maintain Pituitary and Serum Follicle-Stimulating Hormone in Gonadotropin-Releasing Hormone Antagonist Treated Male Rats

Recently it has been found that testosterone can maintain and restimulate serum and pituitary follicle-stimulating hormone (FSH) in the gonadotropin-releasing hormone (GnRH) antagonist treated adult male rat. The present investigation was undertaken to determine (1) which metabolite of testosterone, dihydrotestosterone (DHT), or estradiol accounts for the effects of testosterone in GnRH antagonist suppressed rats and (2) whether these effects of testosterone are influenced by other testicular factors. Eight groups of 6-8 adult male Sprague-Dawley rats were subjected to the following treatments: vehicle, GnRH antagonist (75 micrograms/day s.c.), testosterone-filled Silastic implants (3 x 5 cm, s.c.), DHT-filled Silastic implants (3 x 5 cm, s.c.), estradiol benzoate (15 micrograms/day s.c.), and combined administration of GnRH antagonist with either steroid. In addition, the GnRH antagonist/testosterone treatment regimen was applied to rats orchidectomized 72 h prior to initiation of treatments. After 3 weeks of treatment, serum was analyzed for concentrations of luteinizing-hormone (LH), FSH, testosterone, DHT, and estradiol. Pituitary extracts were analyzed for LH and FSH content. Except for the vehicle-treated groups, serum and pituitary LH concentrations were markedly suppressed by all treatments. In intact rats treated with GnRH antagonist alone and/or estradiol, the pituitary FSH level was reduced by more than 70% relative to controls, while both testosterone and DHT maintained pituitary FSH. Similarly, testosterone and DHT, but not estradiol, delayed the decline of serum FSH induced with GnRH antagonist alone. In orchidectomized animals, testosterone was also capable of preventing a reduction of pituitary FSH despite concomitant GnRH antagonist administration.(ABSTRACT TRUNCATED AT 250 WORDS)

Basal gonadotropin hormone release rates during the period of selective follicle-stimulating hormone release in the juvenile female rat

There are situations in which adult female rats release increased amounts of follicle-stimulating hormone (FSH) independent of increased luteinizing hormone (LH) release. This results from, at least in part, a selective increase in the basal FSH release rate. We investigated whether an increase in the basal FSH release rate is contributory to the rise in serum FSH levels which occurs independent of a rise in serum LH levels in the immature female rat. Rats had high serum FSH concentrations on days 7 and 15 after birth, low serum FSH levels on day 23, and low serum LH levels on all three days. In contrast, anterior pituitary gland (APG) FSH and LH concentrations and contents increased from day 7 to day 15 and the contents increased further from day 15 to day 23. Similarly, basal FSH and LH release rates per mg APG or per APG, as assessed by measurement of FSH and LH released into culture medium containing APG(s) from different aged rats, increased from day 7 to day 15 but did not increase further between days 15 and 23. The results indicate that unlike situations observed to date in adult female rats, a mechanism(s) other than an increase in the basal FSH release rate is involved in selective FSH release in the immature female rat.

Lighting Conditions Affect Testosterone Feedback Sensitivity in Castrated Rats

It has been shown in the Syrian hamster that a short photoperiod sensitizes the hypothalamo-hypophyseal axis of castrated animals to the negative feedback effect of testosterone. There is some evidence that even the reproductive system of the rat, which is generally considered not to be very sensitive to light, can respond to changes in illumination. Therefore, we found it of interest to examine whether alterations in lighting conditions produce changes of sensitivity in the negative feedback effect of testosterone in the rat. We kept intact, castrated, and castrated testosterone-treated animals either in periodic (L:D 12:12) or constant light for 7 days starting 4 weeks after castration. In all 3 testosterone-injected groups, serum luteinizing hormone (LH) was lower in constant than in periodic light. Exogenous testosterone did not decrease the castration-induced elevations of pituitary LH and follicle-stimulating hormone (FSH). On the contrary, testosterone increased the pituitary contents of LH and FSH, especially in constant light. We conclude that, in constant light, the hypothalamo-hypophyseal axis of the castrated rat becomes more sensitive to the negative feedback action of testosterone.

Short-term pituitary-testicular responses of prepubertal ram lambs to hemicastration.

To examine the short-term effects of hemicastration on pituitary-gonadal responses, 12 ram lambs were anesthetized and hemicastrated at 4 mo of age and killed (n = 4) at 2 (HC2), 7 (HC7), or 14 (HC14) days following surgery. Four intact (INT) rams were killed 14 days following anesthesia. Testis and pituitary weights were similar between HC and INT rams. Serum follicle-stimulating hormone (FSH) in HC rams increased within 6 h, peaked at 12 h, and remained elevated above INT levels throughout the study. Overall mean serum testosterone levels in HC rams were lower than in INT rams for the first 48 h, but were similar by 3 days post-surgery. Pulsatile luteinizing hormone (LH) and testosterone secretion was suppressed for the first 9.5 h following anesthesia and/or surgery in both HC and INT animals. A single LH pulse and succeeding testosterone pulse occurred in 10/12 HC and 4/4 INT rams between 10 and 14 h post-surgery, both of which were lower in amplitude in HC than INT animals. However, on Day 7, pulsatile secretory patterns of LH and testosterone were similar, suggesting compensatory androgen secretion had occurred in HC rams. Pituitary LH content was unaffected by hemicastration. In contrast, pituitary FSH content was greater in HC7 and HC14 compared to HC2 and INT animals. Pituitary gonadotropin hormone-releasing hormone (GnRH) receptor concentrations were similar in INT, HC7, and HC14 rams, but were slightly reduced in HC2 rams. Neither testicular LH nor FSH receptor concentrations were altered by hemicastration at any time.(ABSTRACT TRUNCATED AT 250 WORDS)

Pituitary Receptors for Gonadotropin-Releasing Hormone in Relation to Changes in Pituitary and Plasma Gonadotropins in Ovariectom ized Hypothalamo/Pituitary-Disconnected EWES. II. A Marked Rise in Receptor Number during the Acute Feedback Effects of Estradiol1

Ovariectomized (OVX), hypothalamo/pituitary-disconnected (HPD) ewes were used to ascertain the short-term effects of estradiol on the number of gonadotropin-releasing hormone (GnRH) receptors in the pituitary gland. The time course of the study was such that measurements were made during the period of short-term negative feedback and positive feedback. Groups of 4 OVX-HPD ewes were given 250-ng pulses of GnRH each hour and an i.m. injection of oil (Group 1) or 50 micrograms estradiol benzoate in oil (Groups 2-4). Blood samples were collected from each ewe prior to treatment with estradiol or oil and again immediately before slaughter. Groups 2, 3, and 4 were killed 6, 16, and 20 h, respectively, after administration of estradiol. Amplitudes of luteinizing hormone (LH) pulses and average plasma concentrations of LH were reduced 6 h after estradiol treatment. Sixteen and 20 h after injection, the average plasma LH levels were elevated, but pulse amplitudes were similar to preinjection values. The number of GnRH receptors was significantly (p less than 0.01) increased within 6 h of estrogen treatment and further increased 16 and 20 h after treatment. Pituitary content of LH was similar in all groups. These data indicate that the number of GnRH receptors in the pituitary gland of ewes can be acutely influenced by a direct effect of estradiol. However, the magnitude and direction of the change in receptors number does not account for the changes in pituitary responsiveness to GnRH, suggesting estradiol also modifies post-receptor mechanisms that influence secretion of LH.

Pituitary gonadotropin content in gonadectomized rats. Immunoassay measurements influenced by extraction solvent and testosterone replacement.

Increasing interest in events regulating gonadotropin synthesis and secretion led to the reexamination, by radioimmunoassay (RIA), of the pituitary content of luteinizing hormone (LH) and follicle stimulating hormone (FSH). In a preliminary experiment, the importance of the composition of the solvent used to homogenize pituitaries on the detection of LH and FSH was demonstrated by RIA. Experiment 1 demonstrated that a drop in pituitary FSH occurred acutely following gonadectomy in males, but not in females. Experiment 2 tested whether this decline resulted from loss of testosterone or inhibin. Males were castrated and treated daily with oil or testosterone propionate, porcine follicular fluid, porcine serum, or testosterone plus follicular fluid for 2 to 14 days. Castration lowered pituitary FSH at 2, 4, and 6 days. Follicular fluid suppressed serum FSH, but not LH, and did not prevent the fall in pituitary FSH. Testosterone blocked the rise of serum LH and FSH and prevented the decline in pituitary FSH.

Progesterone does not inhibit the increase in pituitary content of luteinizing hormone after removal of estradiol in the ewe.

During late gestation in the ewe, the pituitary content of LH is reduced by about 95%, presumably due to the presence of high concentrations of ovarian steroids. The aim of this study was to determine whether the pituitary content of LH in the ewe can increase after long-term administration of ovarian steroids, when only estradiol (E) is removed or if both E and progesterone (P) must be withdrawn to allow synthesis of LH to occur. Ten ovariectomized ewes were treated with implants containing E and P. After 3 weeks of treatment, the E implants were removed from 5 ewes (-E+P) and both steroid implants were removed from the remaining 5 ewes (-E-P). Five ovariectomized ewes received P implants at the beginning of the experiment and these implants were left in place for the duration of the study; 5 ovariectomized ewes served as controls (C). All animals were injected with 100 micrograms GnRH iv 3, 6 and 9 weeks after the initiation of treatment. The area under the LH-response curve was used as an indication of the pituitary content of LH. All steroid treatments markedly reduced basal levels of LH. LH levels increased only in -E-P ewes, beginning 6 weeks after initiation of the study. After 3 weeks, -E+P and -E-P ewes released less LH (P less than 0.05) in response to GnRH than did C ewes, whereas P animals did not differ from controls. LH release in response to GnRH in -E+P and -E-P groups had increased by 6 and 9 weeks and was not different from that of C ewes.(ABSTRACT TRUNCATED AT 250 WORDS)

Dual role of glucocorticoids in regulation of pituitary content and secretion of gonadotropins

To verify the inhibitory effect of cortisol (F) on secretion of luteinizing hormone (LH) 24 h postorchidectomy, we implanted cholesterol (C) or F subcutaneously into male rats, and 4 days later orchidectomized or sham orchidectomized them under ether anesthesia. We injected gonadotropin-releasing hormone (GnRH) or saline into these rats 24 h postorchidectomy, collected blood 30 min later, and measured LH and follicle-stimulating hormone (FSH) in serum and pituitaries. F inhibited GnRH-induced secretion of LH without affecting secretion of FSH. We then implanted C, corticosterone (B), or F into rats, performed the same surgeries, and collected pituitaries 24 h after surgery for quantitation of receptors for GnRH. Neither F nor B affected the number of receptors for GnRH or their affinity for a GnRH analogue. Suppression of LH in serum occurred without decreased pituitary content of LH. In contrast, F increased pituitary content of FSH. Implantation of progesterone in a similar experiment did not affect circulating concentrations or pituitary contents of FSH or LH. These data suggest that glucocorticoids may inhibit responsiveness to GnRH by some mechanism distal to the receptor for GnRH that affects only LH.

Pituitary gonadotropin-releasing hormone receptor content in rats with polycystic ovaries.

A single injection of estradiol valerate (EV) induces, after a lag period of 4-6 wk, a chronic anovulatory polycystic ovarian (PCO) condition in adult rats. This condition is associated with a selective compromise of luteinizing hormone (LH) release and/or synthesis reflected in low basal serum LH concentrations, decreased pituitary content of LH, and decreased gonadotropin-releasing hormone (GnRH)-stimulated LH secretion. The present study was undertaken to determine to what extent the aberrant LH release in rats with PCO could be related to alterations in pituitary content of GnRH receptors. Pituitary GnRH-receptor content was assessed by the evaluation of saturation binding of a GnRH analog, [125I]-D-Ala6-des-Gly10-GnRH, to pituitary membrane preparations. The receptor content of pituitaries from rats with PCO was compared to that obtained from intact animals at estrus and diestrus. Receptor levels in ovariectomized normal rats and rats with PCO were also assessed. The pituitary GnRH receptor content in PCO rats was similar to that observed in normal controls at estrus and was significantly lower than that for rats at diestrus. Although a twofold increase in pituitary GnRH receptor content was observed at 28 days following the castration of control rats, GnRH receptor content in the pituitaries of PCO rats, at 28 days following ovariectomy, remained unchanged. Although, castration-induced elevations in mean serum LH and follicle-stimulating hormone (FSH) concentrations were observed in both the PCO and control animals, the rise in both gonadotropins was significantly attenuated in the PCO-castrates when compared to the ovariectomized controls. Since GnRH is a major factor in the regulation of pituitary GnRH receptor content, these findings suggest that hypothalamic GnRH release is impaired in rats with PCO and that this impairment is independent of any influences from the polycystic ovaries.

Pituitary receptors for GnRH and estradiol, and pituitary content of gonadotropins in beef cows. II. Changes during the postpartum period

Pregnant beef heifers (n = 24) were assigned randomly to four groups and slaughtered at day 1, 15, 30 or 45 postpartum. The day prior to slaughter blood samples were taken from each cow every 15 min for 8 hr. The anterior pituitary gland, preoptic area (POA) and medial basal hypothalamus (HYP) were collected from each cow. Contents of gonadotropin-releaasing hormone (GnRH) in extracts of POA and HYP, and luteinizing hormone (LH) and follicle-stimulating hormone (FSH) in extracts of anterior pituitary were quantified by radioimmunoassay. In the anterior pituitary gland, membrane receptors for GnRH were quantified by a standard curve technique and cytosolic receptors for estradiol were quantified by saturation analysis. Concentrations of LH, FSH and prolactin in serum were quantified by radioimmunoassay. Only one cow of eight had a pulse of LH during the 8 hr bleeding period on day 1 postpartum. This increased to 8 pulses in 6 cows on day 30 postpartum. Contents of GnRH in POA (15.0 ± 3.2 ng) and HYP (14.0 ± 2.0 ng) did not change significantly during the postpartum period. Pituitary content of LH was low following parturition (.2 ± .1 mg/pituitary) and increased significantly through day 30 postpartum (1.2 ± .1 mg/pituitary). Pituitary content of FSH did not change over the postpartum period. Receptors for both GnRH (.9 ± .2 pmoles/pituitary) and estradiol (5.0 ± .9 /moles/pituitary) were elevated on day 15 postpartum, possibly increasing the sensitivity of the anterior pituitary gland to these hormones and leading to an increased rate of synthesis of LH that restored pituitary content to normal by day 30 postpartum.

LH subunit mRNA concentrations during LH surge in ovariectomized estradiol-replaced rats.

In cycling rats, pituitary concentrations of luteinizing hormone (LH) beta-subunit mRNA increase two- to threefold before the afternoon proestrus LH surge without a corresponding increase in alpha-subunit mRNA. Estradiol (E2) treatment is known to allow expression of daily LH surges in ovariectomized (OVX) rats, and the timing, magnitude, and duration of LH secretion is similar to the LH surge on proestrus. The present study was conducted to examine whether the regulation of LH subunit mRNAs during the LH surge in OVX-E2-treated rats is similar to that present on proestrus. Female Holtzman rats were OVX and Silastic implants containing E2 were inserted subcutaneously under ether anesthesia. Some animals received bromocriptine (0.6 mg sc, twice/day beginning 1 h before surgery). On the 2nd day after surgery, groups of animals (n = 4-10/group) were decapitated at intervals between 1000 and 2100. LH and prolactin (PRL) levels were measured in trunk blood. LH subunit mRNA concentrations in the pituitaries were measured by dot-blot hybridization assay. In OVX-E2 rats the LH surge occurred at 1830 and was accompanied by a selective twofold increase in alpha-subunit mRNA (from 266 +/- 18 to 459 +/- 61 pg cDNA bound/100 micrograms pituitary DNA) and maximum values were present at 1730. LH beta-subunit mRNA (m = 29 +/- 1 pg cDNA bound/100 micrograms pituitary DNA) was unchanged throughout the day. Bromocriptine treatment resulted in the suppression of serum PRL (m = 23 +/- 2 ng/ml) and the LH surge was delayed by 1-1.5 h and somewhat blunted.(ABSTRACT TRUNCATED AT 250 WORDS)