Luteinizing Hormone Synthesis Research Articles

Modulatory actions of estradiol and progesterone on phorbol ester-stimulated LH secretion from cultured rat pituitary cells

We compared the ability of estradiol and progesterone to modulate gonadotropin-releasing hormone (GnRH) and protein kinase C (PKC)-mediated luteinizing hormone (LH) secretion. Long-term (48 h) treatment of rat pituitary cells with 1 nM estradiol enhanced GnRH and phorbol ester (TPA)-stimulated LH secretion. This positive effect was facilitated by additional short-term (4 h) treatment with progesterone (100 nM). However, long-term progesterone treatment, which inhibited GnRH-stimulated LH secretion, did not influence TPA-stimulated gonadotropin release. These steroid actions occurred without an effect on the total amount of LH in the cell cultures (total LH = LH secreted + LH remaining in the cell) and neither the secretagogues nor the steroids altered total LH. Since GnRH or TPA-induced LH secretion depends on Ca 2+ influx into the gonadotroph, we also analyzed the effects of estradiol and progesterone under physiological extracellular Ca 2+ concentrations and in the absence of extracellular Ca 2+. The steroids were able to influence GnRH or TPA-induced LH secretion under both conditions. However, when TPA was used as stimulus in Ca 2+ -deficient medium the relative changes induced by estradiol and progesterone were more pronounced, possibly indicating that the extracellular Ca 2+ -independent component of PKC-mediated LH secretion is more important for the regulation of the steroid effects. It is concluded that estradiol and progesterone might mediate their modulatory actions on GnRH-stimulated LH secretion via an influence on PKC. This effect can occur independently from de novo synthesis of LH and Ca 2+ influx into gonadotrophs.

Luteinizing hormone synthesis in cultured fetal human pituitary cells exposed to gonadotropin-releasing hormone

Luteinizing hormone synthesis in cultured fetal human pituitary cells exposed to gonadotropin-releasing hormone

Luteinizing hormone synthesis in cultured fetal human pituitary cells exposed to gonadotropin-releasing hormone.

To investigate whether GnRH regulates LH synthesis during human development, pituitary cells from second trimester fetuses were incubated with [35S]methionine ([35S]met) and [3H]glucosamine ([3H]gln) for 48 h with 0, 10(-9), and 10(-7) mol/L GnRH. Immunoassayable (i) LH was measured in media and cellular lysates, and dual label scintillation analysis was used to quantitate incorporation of radiolabeled precursors into cells, trichloroacetic acid-precipitable proteins, and immunoprecipitated LH subjected to electrophoresis. Exposure of cells to GnRH did not affect cellular uptake or incorporation of precursors into proteins, but specifically increased total (secreted plus cellular) LH synthesis. Both GnRH concentrations significantly increased iLH release and enhanced secreted and cellular [3H]gln-LH. The secretion of [35S] met-LH was stimulated only by 10(-7) mol/L GnRH. The proportion of newly synthesized LH that was secreted and the 3H/35S ratio of secreted and cellular LH were uninfluenced by GnRH. Although basal LH synthesis was not sex dependent, total iLH content and GnRH-stimulated LH translation were greater in cells from females than in those from males. Therefore, GnRH regulates LH synthesis by second trimester fetal human gonadotrophs without influencing the proportion of total radiolabeled LH that is secreted. The existence of a sex difference in total iLH content and GnRH-stimulated LH translation is consistent with the sexual dimorphism in pituitary LH content occurring during human development.

Modulatory effect of steroid hormones on GnRH-induced LH secretion by cultured rat pituitary cells.

The purpose of the present experiments was to examine the short- and long-term effects of estradiol-17 beta (E2), progesterone (P), and 5 alpha-dihydrotestosterone (DHT), alone and in combination, on the gonadotrophin-releasing hormone (GnRH)-induced luteinizing hormone (LH) secretion, using an ovariectomized rat pituitary cells culture model. After 72 h in steroid-free medium, pituitary cells were further cultured for 24 h in medium with or without E2 (1 nM), P (100 nM), or DHT (10 nM). Cultures were then incubated for 5 h in the absence or presence of 1 nM GnRH with or without steroids. LH was measured in the medium and cell extract by radioimmunoassay. The results show that the steroid hormones exert opposite effects on the release of LH induced by GnRH, which seems to be dependent upon the length of time the pituitary cells have been exposed to the steroids. In fact, short-term (5 h) action of E2 resulted in a partial inhibition (64% of control) of LH release in response to GnRH, while long-term (24 h) exposure enhanced (158%) GnRH-induced LH release. Similar results were obtained with DHT, although the magnitude of the effect was lower than with E2. Conversely, P caused an acute stimulatory action (118%) on the LH released in response to GnRH and a slightly inhibitory effect (90%) after chronic treatment. GnRH-stimulated LH biosynthesis was also influenced by steroid treatment. Significant increases in total (cells plus medium) LH were observed in pituitary cells treated with E2 or DHT.(ABSTRACT TRUNCATED AT 250 WORDS)

In vivo effects of acute EtOH on rat α and β luteinizing hormone gene expression

The suppressive effects of ethanol (EtOH) on the male rodent reproductive axis, especially on serum luteinizing hormone (LH) levels, are well known. In this study we examined, in coordinate fashion, the effects of EtOH on LH secretion and on steady-state levels of the mRNA for the two genes that direct LH synthesis, namely alpha- and beta-LH. A single intraperitoneal (IP) injection of EtOH given to castrated male rats produced a statistically significant fall in serum LH levels at 1.5 (p less than 0.05) and 3 hours (p less than 0.01) after injection compared to saline-injected controls. This effect had dissipated by 24 and 72 hours after treatment. Intrapituitary LH content was significantly increased in EtOH-compared to saline-treated animals 1.5 hours after injection (p less than 0.05) at the same time there was a significant decrease in serum LH. Steady-state levels of beta-LH mRNA were significantly diminished in EtOH-treated animals at 1.5 (p less than 0.05) and 3 hours (p less than 0.001) after injection, but returned to control levels thereafter. Since the half-life of beta-LH mRNA is greater than 8 hours, this fall is most likely due to decreased beta-LH mRNA stability and may also involve decreased beta-LH gene transcription. alpha mRNA levels were unchanged at all time points investigated. These findings were verified in four repetitions of this experiment. These data suggest that the EtOH-induced fall in serum LH is due to impaired secretion of LH and to decreased LH synthesis as indicated by diminished steady-state levels of beta-LH subunit mRNA, secondary mainly to altered mRNA stability.

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.

Progesterone modulation of gonadotropin secretion by dispersed rat pituitary cells in culture. I. Basal and gonadotropin-releasing hormone-stimulated luteinizing hormone release

Dispersed, estradiol-treated, rat pituitary cells were cultured to characterize the influences of a physiologic concentration of progesterone (P, 10 −7 M) on gonadotroph responsiveness to gonadotropin-releasing hormone (GnRH). Acute ( $ ̌ 6 h) P treatment enhanced and chronic ( $ ̆ 12 h) treatment suppressed both basal and GnRH-stimulated luteinizing hormone (LH) release. This modulation took place without any change in intracellular LH stores, indicating that the secretory changes are not attributable to changes in LH synthesis, and were not accompanied by similar alterations in basal or thyrotropin-releasing hormone-stimulated prolactin secretion. Moreover, the timing of these responses was fixed since a 10-fold lower P concentration produced only smaller and briefer alterations in LH release. Analyses of the temporal characteristics of effective P stimuli indicated that a brief 6 h exposure to P inhibited GnRH-stimulated LH secretion 18 h later. In contrast, P's acute actions rapidly dissipated following removal of the steroid from the culture medium. Finally, P-induced enhancement and suppression of GnRH-stimulated LH release could be blocked by appropriately timed treatments with protein synthesis inhibitors. Our findings are consistent with the hypothesis that P influences gonadotroph secretory function via the production of specific proteins.

Effect of dexamethasone on secretion of luteinizing hormone and testosterone in the boar

Eight adult, Yorkshire-Landrace crossbred boars were used to evaluate the effects of the synthetic glucocorticoid, dexamethasone (DXM) on the secretion of luteinizing hormone (LH) and testosterone. Four treatments of 4 d each were administered: 1) 2 ml i.m. of 0.9% (w/v) NaCl solution (control); 2) DXM (2 ml i.m. as a dose of 50 μg/kg body weight, every 12 h); 3) DXM plus gonadotropin releasing hormone (GnRH; 50 μg in 1 ml i.m. every 6 h); 4) 2 ml NaCl solution i.m. plus a single dose of 50 μg i.v. GnRH. Blood samples were collected twice daily from an indwelling jugular vein catheter for 3 d and at 15 min intervals for 12 h on the fourth day. DXM treatment resulted in lower (P M0.01) testosterone values in samples collected twice daily. More frequent sampling on Day 4 revealed that DXM reduced (P<0.01) the number of pulsatile increases of LH in plasma, although the individual mean pulse areas did not fiffer between the NaCl- and DXM-treated groups. This was associated with a decreased pulse frequency of testosterone (P<0.05). GnRH plus DXM treatment caused a significant elevation (P<0.05) in mean values as well as in the mean pulse area and in the total of the individual pulse areas of LH. Pulse area and mean concentrations of testosterone were also increased (P<0.01) when GnRH was given concurrently with DXM. Comparison of a single injection of GnRH when NaCl was being administered (Treatment 4) to one of the injections of GnRH (Day 4, 0800 h, Treatment 3) revealed a subsequently greater (P<0.01) pulse area in LH above base-line during DXM treatment (7.67 ± 1.17 ng/ml) than during the NaCl (4.17 ± 0.73 ng/ml) treatment period. This was reflected in a greater (P<0.01) pulse increase of testosterone following the LH pulse in boars treated with DXM. It is concluded that DXM treatment in the boar can reduce the pulse frequency of LH secretion, presumably by affecting GnRH secretion, but it has less effect directly on pituitary LH synthesis and release.

Expression of α-Subunit and Luteinizing Hormone (LH) β Messenger Ribonucleic Acid in the Rat during Lactation and after Pup Removal: Relationship to Pituitary Gonadotropin-Releasing Hormone Receptors and Pulsatile LH Secretion*

Pituitary GnRH receptor (GnRH-R) levels and LH secretion are suppressed in the lactating rat. To determine if LH synthesis is also inhibited, we have measured LH subunit mRNA levels in the pituitary of lactating rats. We have also examined the temporal relationship among restoration of GnRH-R, LH secretion, and LH synthesis after withdrawing the sensory stimulus of suckling. Pituitary alpha-subunit and LH beta mRNA levels were sharply reduced on day 10 of lactation in both intact and ovariectomized (OVX) animals compared with those in cycling diestrous rats or OVX controls. Removal of the suckling stimulus from OVX animals led to significant increases in alpha-subunit and LH beta mRNA levels by 24 h. Upon removal of the suckling stimulus from intact rats, alpha-subunit mRNA levels were restored by 48 h, but LH beta mRNA levels did not return to diestrous levels until 72 h. Pituitary GnRH-R levels were clearly up-regulated within 1 day after pup removal. Some LH pulses were observed by 48 h, but consistent plasma LH pulses were not detected until 72 h. When pulsatile GnRH was administered during the 24 h after pup removal from intact rats, the regimen of pulsatile GnRH was successful in inducing LH secretion; however, the restoration of pulsatile LH was not accompanied by increases in alpha-subunit and LH beta mRNA levels. The present studies provide further evidence to support the hypothesis that during lactation, the suppression of pituitary gonadotroph function is mainly due to the loss of hypothalamic GnRH secretion. Our data also show that 1) the restoration of GnRH-R alone is not sufficient to activate LH subunit mRNA and LH secretion; 2) the normal restoration of pulsatile LH secretion and increases in LH subunit mRNA are temporally correlated, as increases in LH secretion appear to precede increases in LH subunit mRNA; and 3) the restoration of pituitary LH subunit mRNA levels and pulsatile LH secretion took longer in the intact rat than in the OVX rat, suggesting that ovarian steroids may play a role in the inhibitory effect of lactation.

Evidence for a role of protein kinase C in luteinizing hormone synthesis and secretion. Impaired responses to gonadotropin-releasing hormone in protein kinase C-depleted pituitary cells.

The role of protein kinase C in luteinizing hormone (LH) release was analyzed in studies on the actions of phorbol esters and gonadotropin-releasing hormone (GnRH) in normal and protein kinase C (Ca2+/phospholipid-dependent enzyme)-depleted pituitary cell cultures. LH secretory responses of normal pituitary cells to GnRH were reduced but not abolished in Ca2+-deficient medium, consistent with the existence of extracellular Ca2+-dependent and -independent components of GnRH action. Both of these components could be elicited by treatment with 12-O-tetradecanoylphorbol 13-acetate (TPA). The LH secretory responses to TPA and GnRH were additive only at low doses and converged to a common maximum at high concentrations of the agonists in the presence or absence of extracellular Ca2+. The release of stored LH by GnRH and TPA was accompanied by secretion of newly synthesized LH from 2 to 5 h during stimulation by either of the agonists. LH synthesis was increased in a progressive and dose-dependent manner by GnRH and TPA, and the ratio between newly synthesized and released hormone was near 1:2. TPA caused rapid and complete translocation of cytosolic protein kinase C to the particulate fraction of pituitary cells, followed by a progressive decrease in total enzyme content to approximately 10% after 6 h. Partial recovery of the cytosolic enzyme (to 20%) occurred after washing and reincubation for 15 h. Such kinase C-depleted cells showed prominent, dose-dependent reductions in the actions of GnRH and TPA on LH release and synthesis in both normal and Ca2+-deficient media. These observations support the hypothesis that protein kinase C participates in LH biosynthesis and secretion in pituitary gonadotrophs and is involved in the actions of GnRH upon these processes.

Evidence Suggesting the Trophoblastic Origin of Follicle‐Stimulating Hormone, Luteinizing Hormone and Prolactin in Hydatidiform Mole

AbstractOur previous finding that the intravesicular fluid of hydatidiform mole contains high concentrations of follicle stimulating hormone (FSH), luteinizing hormone (LH) and prolactin (PRL) suggested to us that the molar trophoblast may be able to synthesize polypeptide hormones just as it produces human chorionic gonadotrophin (hCG). Using radioimmunoassay, we examined the plasma concentrations of these hormones before and after evacuation of the molar tissues, because we believed that this procedure might identify the source of increased FSH, LH and PRL synthesis in hydatidiform moles. Our 4 subjects had markedly elevated pre‐evacuation plasma levels of FSH (9.0–12.5 IU/l), LH (135,000–340,000 IU/l), PRL (1,300–3,800 mIU/l) and β‐hCG (550,000–4,500,000 IU/l). Their liver and kidney function tests were normal. After molar evacuation, serial concentrations of FSH, LH and PRL progressively declined to near normal levels by 9–16 weeks. As expected, the changes in FSH, LH and PRL levels closely resembled the typical pattern of changes in β‐hCG levels and decreased to the lower limits of sensitivity of the assay (&lt;2.5 IU/l at 9–12 weeks). The pattern and magnitude of the changes strongly suggest that the diseased trophoblast makes a significant contribution to circulating levels of the 4 hormones.

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.

An intractable, ovary-independent impairment in hypothalamo-pituitary function in the estradiol-valerate-induced polycystic ovarian condition in the rat.

A single injection of estradiol valerate (EV) produces anovulatory acyclicity and polycystic ovaries (PCO) in the rat. Basal serum luteinizing hormone (LH) concentrations are attenuated whereas serum follicle stimulating-hormone (FSH) concentrations are in the high normal range in these animals. Subsequent unilateral ovariectomy restores ovulatory cycles and normal histology in the remaining ovary without correcting the aberrant basal serum gonadotropin concentrations. This suggests that although the blocked surge mechanism is correctable, a second relatively intractable, ovary-independent impairment compromises basal gonadotropin production. To identify and characterize this second component, we have examined hypothalamic-pituitary function in PCO rats after bilateral ovariectomy. Adult (200-250 g), normal cyclic Wistar rats were injected with 2 mg EV or with vehicle (control). Nine weeks later all animals were ovariectomized and PCO was confirmed in the EV-treated animals. Animals were killed at 0, 2, 7, 14, and 28 days after ovariectomy, and hypothalamic content of luteinizing hormone-releasing hormone (LHRH) and pituitary and serum concentrations of LH and FSH were measured. LH and FSH responses to exogenous LHRH were assessed. Serum progesterone, testosterone, and estradiol concentrations were determined at 28 days. Hypothalamic LHRH decreased significantly in all animals over the 28-day period. Although LHRH values did not differ at Time 0, by 28 days there was significantly less LHRH in the hypothalami of control than in PCO rats. This pattern of depletion was mirrored by corresponding reciprocal patterns of increasing serum gonadotropin concentrations.(ABSTRACT TRUNCATED AT 250 WORDS)

Pituitary receptors for GnRH and estradiol, and pituitary content of gonadotropins in beef cows. I. Changes during the estrous cycle

To further characterize the endocrinological changes in the hypothalamo-hypophyseal axis thoughout the bovine estrous cycle, cycling beef heifers (n=24) were randomly assigned to six groups. These heifers were slaughtered 6, 12, 18, 19, 20 or 21 days following their previous estrus (day 0). Anterior pituitaries and hypothalami were collected. Hypothalami were divided into the preoptic area and medial basal hypothalamus, and content of gonadotropin-releasing hormone (GnRH) was quantified by radioimmunoassay. Contents of luteinizing hormone (LH) and follicle stimulating hormone (FSH) in the anterior pituitary gland were quantified by radioimmunoassay. Membrane receptors for GnRH were quantified by a standard curve technique and receptors for estradiol in anterior pituitary cytosol were quantified by saturation analysis. There was no significant change in content of GnRH in the hypothalamus or content of FSH in the anterior pituitary on any of the days examined; however, content of GnRH in the preoptic area was lower (P<.1) on day 19 postestrus. Cytosolic receptors for estradiol increased (P<.05) on day 18 post-estrus and returned to baseline by day 19. Content of LH and the number of receptors for GnRH in the anterior pituitary gland decreased (P<.01) on day 19 postestrus, and the number of receptors for GnRH remained low through day 21 postestrus. The reduction in anterior pituitary content of LH was transient indicating that synthesis of LH restores pituitary content to preovulatory levels before the number of receptors for GnRH returns to normal.

Selective modulation of FSH and LH secretion by steroids.

Significant divergence between the pattern of FSH and LH secretion has been observed in the ovulatory cycle, after ovariectomy and during puberty. The presence of an FSH-releasing factor, gonadal FSH inhibiting and releasing peptides and changes in the pulsatile pattern of LHRH secretion are among the postulates used to explain the divergent secretion of FSH and LH. Experiments in our laboratory have shown considerable evidence of differential regulation of FSH and LH secretion by steroids in the absence of gonadal regulatory peptides. Natural and synthetic estrogens show significant differences in the suppression of FSH and LH in the ovariectomized rat using a standard uterine response to the estrogen as the end point. In the immature ovariectomized rat treated with a low dose of estradiol that is sufficient for the synthesis of progesterone receptors to ensure progesterone sensitivity, but not large enough to induce estrogen triggered LH surges, progesterone administration resulted in a pattern of LH and FSH secretion similar to that observed on the day of proestrus in the cycling rat. Selective secretion of FSH was induced in the estrogen primed immature rat model by the administration of progesterone metabolite 5 alpha-dihydroporgesterone (5 alpha-DHP) while selective LH secretion was induced by 3 alpha, 5 alpha-tetrahydroprogesterone (3 alpha,5 alpha-THP). The selective secretion of FSH and LH induced by progesterone metabolites was confirmed in the immature female rat primed with PMSG and maintained in constant light. 5 alpha-DHP was also able to induce a greater release of FSH when administered to the adult cycling rat on proestrus. The priming of the pituitary gonadotrope in secreting a high baseline level of FSH or responding to LHRH in releasing a greater amount of FSH appeared to be an important factor in selective FSH release and such priming can be brought about by 5 alpha-DHP in the absence of gonadal regulatory peptides.

Differential effect of glucocorticoids on synthesis and secretion of Luteinizing Hormone (LH) and Follicle Stimulating Hormone (FSH)

Our previous work has suggested that glucocorticoid pretreatment suppresses the enhanced responsiveness to GnRH seen in serum LH 12 h after castration. By contrast, serum FSH continues to show the castration-induced hypersensitivity to GnRH. Our attempts to replicate this LH suppression in static pituitary culture in vitro were not successful. This suggested to us the possibility that corticoids in vivo might be preventing castration-induced increases in pituitary GnRH receptor levels. We tested this at 24 h post-castration and, in fact, corticoids did not suppress the increase in GnRH receptors. In addition to the aforementioned effects of corticoids, we have seen that cortisol reverses the castration-induced drop in pituitary FSH content. It does this for 7 days post-castration, even though it no longer has an effect in suppressing serum LH. Thus, our accumulated data reveal that glucocorticoids have a differential effect on LH and FSH synthesis and secretion. Further studies are needed to clarify the site(s) of action of glucocorticoids in gonadotropin secretion and synthesis. Glucocorticoids may well prove to be a key in unlocking the mystery of the mechanism of differential control of regulation of LH and FSH.

Effects of castration or testosterone implants upon pituitary function in hypogonadal mice bearing normal foetal preoptic area grafts.

Grafts of normal mouse preoptic area (POA) tissue into the third ventricle of gonadotrophin-releasing hormone (GnRH)-deficient hypogonadal (hpg) mice resulted in an elevation of pituitary GnRH receptors, an increased synthesis of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) by the pituitary gland, an elevation of gonadal LH receptors and in the stimulation of steroidogenesis and spermatogenesis in the testis. In normal mice both castration or the subcutaneous implantation of testosterone capsules for 10 days reduced GnRH receptors, pituitary LH and FSH content, and the latter treatment also caused a 50% reduction in testicular LH receptors. In hpg mice bearing POA grafts testosterone implants failed to affect any of the above parameters, and castration failed to affect pituitary gonadotrophin hormone content, although there was a slight reduction in pituitary GnRH receptors after castration. These experiments suggest that neither the pituitary gonadotroph, nor the GnRH neurone represent major sites for the direct negative feedback of testosterone upon gonadotrophic hormone secretion in male mice.

An in vitro study of LH release, synthesis and heterogeneity in pituitaries from proestrous and short-term ovariectomized rats.

It is known that acute ovariectomy (OVX) greatly attenuates the pituitary luteinizing hormone (LH) response to gonadotropin-releasing hormone (GnRH) in vitro. The present study evaluated possible quantitative and/or qualitative differences in the biosynthesis and secretion of LH in pituitaries from proestrous and acutely (72 h) OVX rats. Paired anterior pituitary glands were incubated for 4 h in a medium containing +/- 10 nM GnRH. Pituitary and secreted LH were measured by radioimmunoassay with differences in total LH (tissue plus medium) +/- GnRH being indicative of GnRH-stimulated LH synthesis. Qualitative changes in LH were evaluated by isoelectrofocusing (IEF). The results show that the major form of LH stored in and released from the pituitaries consisted of LH molecules with an isoelectric point (pI) in the alkaline pH range (alkaline LH), and a lesser amount (approximately 30%) of LH molecules in the acidic pH range (acidic LH). The ratio of alkaline/acidic LH observed in the pituitary and medium was similar in the proestrous and OVX groups, although the amount of alkaline and acidic LH release in response to GnRH was 2-3 times greater in the proestrous group. In both groups, the alkaline/acidic LH ratio of secreted LH was higher in the presence of GnRH than in its absence. Alkaline LH synthesis was increased by GnRH in both groups, with the response being greater in the proestrous than in the OVX group; GnRH-stimulated acidic LH synthesis was observed only in the proestrous group. In both groups, the amount of LH synthesized was about 60% of the amount released, which suggests that LH synthesis does not fully account for differences in GnRH-stimulated LH release. Treatment of pituitary extracts with neuraminidase decreased acidic LH, and proportionately increased alkaline LH. These results suggest that the quality of LH stored in and secreted from pituitaries of proestrous and OVX rats is similar, and that there is a preferential release of the major alkaline LH isoform in response to GnRH. The ovarian steroid environment, presumably estradiol, proportionately increases the amount of alkaline and acidic LH released, and differentially affects the amounts of the various isoforms synthesized in response to GnRH. The charge heterogeneity of alkaline and acidic LH may be related to the sialic acid content of the LH molecule.

Functional changes in luteinizing hormone-secreting cells from pre- and postpartum ewes.

Luteinizing hormone (LH)-containing cells from ovine pituitaries obtained during gestation and at various times after parturition were examined to determine whether the ability to store and secrete LH in vitro was correlated with morphological changes. Pituitaries collected on days 50 and 140 of gestation and on days 2, 13, 22, and 35 after parturition were enzymatically dissociated and the resulting cells cultured in media containing estradiol (12 pg/ml), cortisol (12 ng/ml), or no steroid. After 4, 7, or 10 days of culture, cells were washed and basal LH, gonadotropin-releasing hormone (GnRH)-stimulated release of LH, and cellular content of LH were determined. The content of LH (ng/10(6) cells) was lowest on day 140 of gestation (2.7 +/- 0.3) and day 2 postpartum (2.2 +/- 0.6) and then increased (P less than 0.05) on days 13 (36.6 +/- 8.3), 22 (59.9 +/- 14.4), and 35 (54.6 +/- 19.3) postpartum. The percentage of pituitary cells containing immunoreactive LH nearly doubled (P less than 0.05) between days 2 (5.6 +/- 0.2%) and 35 (10.6 +/- 1.1%) postpartum. Moreover, LH-containing cells were smaller, and the percent total cellular volume occupied by secretory granules was less on day 2 than on days 22 and 35 after parturition. Secretion of LH after 4, 7, or 10 days of culture reflected the cellular content of LH and was not influenced by the presence of steroids in the media. These data indicate that decreased synthesis of LH during gestation is associated with hypoplasia of the LH-secreting cells. These cells are reactivated during the postpartum period and their capacity to synthesize LH gradually returns to normal.

Immunohistochemical localization of follicle-stimulating hormone, luteinizing hormone, growth hormone, adrenocorticotrophic hormone and prolactin in the human placenta

The sites of localization of luteinizing hormone (LH), follicle-stimulating hormone (FSH), growth hormone (GH), adrenocorticotrophic hormone (ACTH) and prolactin (PRL) within placental tissues have been studied by an immunoperoxidase technique. The syncytiotrophoblast is the sole significant site of localization of LH, FSH, GH and ACTH; PRL is found both in syncytiotrophoblast and in decidual cells. It is highly probable that the sites of localization of these peptide hormones represents their sites of synthesis in the placenta and thus that the syncytiotrophoblast is the sole site of synthesis of LH, FSH, LH and ACTH. PRL appears to be synthesized both in syncytiotrophoblast and decidua, but the latter is probably not the major site of synthesis of this hormone. Whether these placental peptide hormones have any physiological role to play during pregnancy or whether the placental capacity to synthesize such hormones is an atavistic phenomenon of no functional importance is currently a moot point.