Kinetic evidence for two components in the priming effect of LH-releasing hormone in the rat

1991 ◽  
Vol 129 (3) ◽  
pp. 351-355 ◽  
Author(s):  
M. S. Johnson ◽  
R. Mitchell
Keyword(s):  
Priming Effect ◽  
Maximum Amount ◽  
Response Curves ◽  
Initial Exposure ◽  
Low Concentrations ◽  
Before And After ◽  
Lh Release ◽  
Stimulus Secretion Coupling ◽  
Lh Releasing Hormone

ABSTRACT The priming effect of LHRH on LH release from prooestrous rat hemipituitary glands in vitro was analysed by kinetic approaches. Concentration–response curves for LHRH-, K+- and ionomycin-induced LH release were constructed for initial exposure to the secretagogues and after 'priming' with a low dose of LHRH (100 pg/ml). These data were analysed by a non-linear curve-fitting programme to reveal the potency and maxima of the responses before and after priming. The parameters obtained from the curves fitted to the LHRH concentration–response curves showed that two changes had occurred as a result of priming. There was an increase in the maximum amount of hormone released and also a relatively greater ability for low concentrations of LHRH to cause release (increased potency). The data for K+ and ionomycin revealed only one change as a result of priming, an increase in the maximum amount of hormone available for release. The data indicate that LHRH, after self-priming, releases more hormone by at least two routes, one represented by a general increase in stimulus–secretion coupling (which is available to K+ and ionomycin), the other a specific up-regulation of signal transduction by the LHRH receptor–effector system. Journal of Endocrinology (1991) 129, 351–355

Changes in the granule population of gonadotrophs of hypogonadal (hpg) and normal female mice associated with the priming effect of LH-releasing hormone in vitro

1986 ◽  
Vol 109 (1) ◽  
pp. 35-NP ◽  
Author(s):  
C. E. Lewis ◽  
J. F. Morris ◽  
G. Fink ◽  
M. Johnson
Keyword(s):  
Priming Effect ◽  
Marginal Zone ◽  
Secretory Granules ◽  
Initial Exposure ◽  
Releasing Hormone ◽  
Female Mice ◽  
Mean Diameter ◽  
Lh Release ◽  
The Mean ◽  
Lh Releasing Hormone

ABSTRACT Changes in the size and position of secretory granules in pituitary gonadotrophs have been studied in relationship to LH release and self-priming induced by LH-releasing hormone (LHRH) in pituitary glands from normal and hypogonadal (hpg) female mice. Hemipituitary glands were preincubated and then incubated for either 1 or 2 h in the absence or presence of LHRH (8·5 nmol/l). The glands were either processed for ultrastructural morphometry or homogenized for the determination of pituitary LH content. Morphometry was carried out on gonadotrophs identified by immunocytochemistry for LHβ using the thin/semi-thin section method. Pituitary LH content and the amount of LH released were determined by radioimmunoassay. The amount of LH released in response to the first and second hours of incubation with LHRH were similar in hpg and normal mice with a clear priming effect (three- to fourfold increase in pituitary responsiveness to LHRH) occurring in both strains. Despite a substantially reduced total number of granules (and amount of LH) in unstimulated hpg gonadotrophs, the number of granules in the outer 500 nm marginal zone of the cells was similar to that in normal mice. This could explain the similar amount of LH released from normal and hpg glands by the first LHRH challenge. The initial exposure to LHRH was also associated with a marked translocation of secretory granules from the central to the outer marginal region of cytoplasm subjacent to the gonadotroph plasmalemma, such that in 'primed' glands 60% of granules were found in this marginal zone compared with 40% (hpg) or 33% (normal) in unstimulated glands. The mean diameter of granules in the marginal zone was significantly less than that of granules in the central zone of the gonadotrophs of unstimulated glands from both normal and hpg animals. Exposure to LHRH for 1 h was associated with an increase in the number of small granules in the marginal zone and a significant decrease in the mean diameter of the gonadotroph granule population as a whole. After the primed release of LH, increased proportions of granules were still located in the marginal zone of gonadotrophs, indicating that granule migration continued during the second hour of exposure to LHRH in which primed release occurred. The primed release was associated with a detectable reduction in both the LH and granule content of gonadotrophs in normal, but not hpg glands. The ultrastructural correlates of LH release and LHRH priming were similar in the two strains of mice, and therefore in mice neither the releasing nor the priming effect of LHRH depends upon previous exposure of the pituitary gland to LHRH or ovarian factors. The priming effect was associated with a marked shift of granules towards the plasmalemma and a decrease in granule size which most likely resulted from increased post-translational processing within secretory granules. J. Endocr. (1986) 109, 35–44

Ionophore A23187 partially reverses LH secretory defect of pituitary cells from old rats

1987 ◽  
Vol 253 (3) ◽  
pp. E233-E237
Author(s):  
R. S. Chuknyiska ◽  
M. R. Blackman ◽  
G. S. Roth
Keyword(s):  
Primary Cultures ◽  
Extracellular Calcium ◽  
Base Line ◽  
Ionophore A23187 ◽  
Pituitary Cells ◽  
Lh Release ◽  
Old Rats ◽  
Lh Releasing Hormone ◽  
Lh Secretion

We measured in vitro release of luteinizing hormone (LH) in the presence of 1.5 mM extracellular calcium, with and without LH-releasing hormone (LHRH; 10(-10) to 10(-7) M) or the ionophore A23187 (10(-7) to 10(-4) M), in primary cultures of anterior pituitary cells from intact mature (6 mo) and old (24 mo) male and intact and ovariectomized mature and old female Wistar rats. Base-line as well as LHRH- and A23187-mediated LH secretion was decreased from cells of old rats. However, exposure to A23187 led to a nearly twofold greater augmentation of LH release from cells of old rats, thus decreasing the apparent age-related LH secretory deficit by approximately one-half. We then measured LHRH-mediated (10(-8) M) vs. A23187-mediated (10(-4) M) LH release with and without extracellular calcium (0.08-1.5 mM). For cells from both mature and old rats, there was a similar calcium dependency for A23187- and LHRH-mediated LH release, with optimal LH secretion at 1.0-1.5 mM extracellular calcium concentrations. Again, both LHRH- and A23187-stimulated LH release was significantly lower and exposure to A23187 led to a greater increase in LH release from cells of old rats. Taken together with similar findings in other systems, these data suggest that the in vitro LH secretory defect of pituitary cells from old rats results in part from one or more defects in calcium mobilization and that such alterations may be a widespread manifestation of aging.

scholarly journals Regulation of gonadotrophin secretion by inhibin, testosterone and gonadotrophin-releasing hormone in pituitary cell cultures of male monkeys

1998 ◽  
Vol 159 (1) ◽  
pp. 103-110 ◽  
Author(s):  
U Fingscheidt ◽  
GF Weinbauer ◽  
HL Fehm ◽  
E Nieschlag
Keyword(s):  
Dose Response ◽  
Pituitary Cells ◽  
Basal Secretion ◽  
Response Curves ◽  
Gonadotrophin Secretion ◽  
Lh Release ◽  
Gonadotrophin Releasing Hormone ◽  
Dose Response Curves ◽  
Male Rhesus

The effects of bovine inhibin, testosterone and GnRH on gonadotrophin secretion by primate pituitary cells were characterized in vitro using pituitaries from six male rhesus monkeys and one male cynomolgus monkey. The effect of inhibin on basal secretion of FSH and LH was investigated. Dose-response curves in monkeys and rats were compared. GnRH dose-response curves in the presence and absence of testosterone were also examined in monkeys. In monkey pituitary cells, testosterone at a concentration of 10(-7) M had no effect on LH or FSH secretion. Inhibin suppressed FSH secretion to 50.8% of that of controls with no effect on LH. In rats, FSH secretion was suppressed to 45.0% of that of controls with a median effective dose (ED50, 95% range) of 1.298 (1.064-1.584) U/ml, compared with 1.024 (0.7204-1.455) U/ml in monkeys. In monkey pituitary cells, LH release was stimulated 9.9-fold and FSH 3.3-fold by GnRH. Testosterone had no effect on basal or GnRH-stimulated gonadotrophin release. These results support the view that the pituitary is not the target organ for the negative feedback action of testosterone in the male. In vitro, inhibin is the major regulator of FSH secretion at the pituitary level.

Evidence for a role of phospholipase A2 in the mechanism of LHRH priming in rat anterior pituitary tissue

1994 ◽  
Vol 141 (1) ◽  
pp. 15-31 ◽  
Author(s):  
F J Thomson ◽  
M S Johnson ◽  
R Mitchell ◽  
B Wolbers
Keyword(s):  
Protein Synthesis ◽  
Anterior Pituitary ◽  
Priming Effect ◽  
Protein Synthesis Inhibitor ◽  
Synthesis Inhibitor ◽  
Pituitary Tissue ◽  
Gonadotrophin Secretion ◽  
Lh Release ◽  
Functional Relevance

Abstract The phospholipase A2 (PLA2) inhibitors, quinacrine, p-bromophenacyl bromide, ONO-RS-082, aristolochic acid and chloracysine blocked the priming effect of LHRH, but not acute LHRH-induced gonadotrophin release measured in anterior pituitary pieces in pro-oestrous rats in vitro. These results suggest that the intracellular mechanisms underlying LHRH priming are distinct from those which mediate LH release in the present circumstances in that they involve PLA2. Furthermore, neither LHRH-induced LH release from preprimed tissue nor Ca2+-induced LH release were attenuated by quinacrine, indicating that this inhibitor does not interfere with the general Ca2+-dependent secretory apparatus of the gonadotroph and that the critical period for its action is in the induction of priming. LHRH induced the release of [3H]arachidonic acid ([3H]AA) from [3H]AA-prelabelled anterior pituitary tissue from pro-oestrous rats; a response which was sensitive to inhibitors of PLA2, of protein kinase C (PKC) and of protein synthesis. Activation of PKC also resulted in [3H]AA release which was inhibited with exactly the same pharmacological profile as the response to LHRH. Both gonadotrophin secretion and [3H]AA release responses to LHRH and to phorbol ester varied in parallel during the oestrous cycle and in ovariectomized/oestradiol-17β-replaced animals, as did their sensitivity to quinacrine and the protein synthesis inhibitor cycloheximide. These results indicate that LHRH priming is dependent on a hormonally regulated cascade involving a distinct form of PKC acting through a protein synthesis-dependent step to release AA by means of PLA2 activity. The priming effect was mimicked (at least in part) by conditioning preincubation with AA, confirming the functional relevance of this signalling cascade. Results using standard inhibitors of lipoxygenase/epoxygenase pathways were equivocal as to whether these pathways were critically involved, whilst cyclo-oxygenase inhibitors were completely without effect. The steps downstream from AA (and its possible metabolites) by which stimulus–secretion coupling is up-regulated in priming remain to be clarified. Journal of Endocrinology (1994) 141, 15–31

The effect of gonadotrophin surge-inhibiting factor on the self-priming action of gonadotrophin-releasing hormone in female rats in vitro

1992 ◽  
Vol 134 (3) ◽  
pp. 427-436 ◽  
Author(s):  
D. W. Koppenaal ◽  
A. M. I. Tijssen ◽  
J. de Koning
Keyword(s):  
Protein Synthesis ◽  
Priming Effect ◽  
The Self ◽  
Pituitary Cells ◽  
Inhibiting Factor ◽  
Lh Release ◽  
Gonadotrophin Releasing Hormone ◽  
Functional Antagonism

ABSTRACT The present study was designed to explore further the functional antagonism between gonadotrophin-releasing hormone (GnRH) and the ovarian factor, gonadotrophin surge-inhibiting factor (GnSIF). In all experiments, pituitary tissue was exposed to various amounts of GnSIF, after which the self-priming action of GnRH was studied. GnSIF was increased in vivo by FSH treatment and increased in vitro by adding various amounts of follicular fluid (FF) to cultured pituitary cells. Treatment with 3 or 10 IU FSH suppressed the initial LH response and delayed the maximally primed LH response to GnRH. Treatment with FSH was only effective in intact rats on days 1 and 2 of dioestrus. There was no difference in the rate of maximal LH release irrespective of treatment with either FSH or saline. Since FSH treatment was ineffective in long-term ovariectomized rats, it was concluded that the initial suppressive effect of FSH on LH release was mediated by GnSIF. Cycloheximide prevented the self-priming action of GnRH by inhibiting GnRH-induced protein synthesis. The initial protein synthesis-independent GnRH-stimulated LH release, which was already suppressed by FSH treatment, remained suppressed in the presence of cycloheximide. Pretreatment with GnRH in vivo increased the protein synthesis-independent GnRH-induced LH release during subsequent incubation of the glands. This increase did not occur after FSH treatment. Pituitary cells, cultured for 20 h in medium only, failed to elicit the self-priming effect of GnRH. Preincubation with FF maintained the self-priming effect. This was independent of the concomitant presence of various amounts of oestradiol. Preincubation with bovine FF suppressed the initial GnRH-stimulated LH release dose-dependently. Porcine FF, human FF and testicular extract suppressed the release of LH in a similar way. It was concluded that GnSIF suppresses the initial LH response to continuous GnRH stimulation. Increased levels of GnSIF caused by FSH treatment also delayed the primed LH release. The mechanism of functional antagonism between GnSIF and GnRH could give rise to the occurrence of the phenomenon of GnRH self-priming. Journal of Endocrinology (1992) 134, 427–436

Estradiol stimulation of LH response to LHRH and LHRH binding in pituitary cultures

1982 ◽  
Vol 242 (6) ◽  
pp. E392-E397
Author(s):  
L. K. Tang ◽  
A. C. Martellock ◽  
J. K. Horiuchi
Keyword(s):  
Cell Proliferation ◽  
Luteinizing Hormone ◽  
Priming Effect ◽  
Cultured Cells ◽  
Female Rats ◽  
Releasing Hormone ◽  
Lh Release ◽  
Estradiol Stimulation ◽  
Lh Releasing Hormone ◽  
Stimulation Of

The relationship between 17 beta-estradiol (E2) stimulation of luteinizing hormone (LH) response to LH-releasing hormone (LHRH) and E2 effect on LHRH binding was examined in pituitary monolayer cultures prepared from female rats. E2 pretreatment significantly (P less than 0.05) augmented the LHRH-induced LH release to 158-180% of the non-E2-treated controls. The maximal E2-priming effect could be observed after 1 day of treatment. E2 treatment for 3 days stimulated [D-Ala6]luteinizing hormone-releasing hormone (LHRHa) binding to about 1.5-fold that of the non-E2-treated controls without affecting the dissociation constant of LHRH receptor (Kd = 4 X 10(-10) M). The stimulatory effect of E2 on cell proliferation as determined by [3H]thymidine incorporation was also observed 3 days after treatment. However, E2 stimulation of LH accumulation in the cultured cells could be detected as early as 4 h after treatment. These results indicate that E2-priming effect on pituitary LH response to LHRH is initially associated with an increase in cellular LH content and later associated with increases in LHRH binding and in an index of cell proliferation that may include the LH-producing cells.

RELEASE OF THYROTROPHIN IN VIVO AND IN VITRO BY SYNTHETIC NEUROHYPOPHYSIAL HORMONES

1964 ◽  
Vol 42 (1) ◽  
pp. 75-83 ◽  
Author(s):  
Frank S. LaBella
Keyword(s):  
Dose Response ◽  
In Vitro Release ◽  
Optimal Level ◽  
Response Curves ◽  
Low Concentrations ◽  
Pituitary Glands ◽  
In Vivo Effects ◽  
Synthetic Oxytocin

The influence of synthetic oxytocin and synthetic lysine-8-vasopressin on the release of thyrotrophin (TSH) from slices of the "basophilic" zone of bovine anterior pituitary glands was determined. Up to 10-fold stimulation of TSH release occurred in the presence of the peptide hormones at low concentrations (approximately 10−11 to 10−9 M). Concentrations greater than 10−9 M were less stimulatory, ineffective, or inhibitory. In general, vasopressin stimulated at lower concentrations than did oxytocin. The dose–response curve of oxytocin began to descend at lower concentrations than did that of vasopressin.Stimulation of I131 discharge from the thyroids of propylthiouracil (PTU)-treated, day-old chicks was produced by the intraperitoneal injection of as little as 4 ng vasopressin or 25 ng oxytocin. As the injected dose of either peptide was increased beyond an optimal level, there was less enhancement of I131 discharge, and, with further increases, inhibition. The decreasing response began with lower doses of oxytocin than of vasopressin. The similarities of the dose–response curves of thyroid I131 discharge and of in vitro release of TSH indicate that the in vivo effects of injected neurohypophysial peptides are mediated through the release of endogenous TSH.

Effects of chronic treatment with oestrogen, an oestrogen antagonist and a potent luteinizing hormone releasing hormone agonist analogue on pituitary responsiveness to luteinizing hormone releasing hormone in the rat

1983 ◽  
Vol 98 (3) ◽  
pp. 313-321 ◽  
Author(s):  
J. M. Hall ◽  
S. A. Whitehead
Keyword(s):  
Luteinizing Hormone ◽  
Chronic Treatment ◽  
Ex Vivo ◽  
Significant Rise ◽  
Luteinizing Hormone Releasing Hormone ◽  
Releasing Hormone ◽  
Serum Lh ◽  
Lh Release ◽  
Lh Releasing Hormone

The rise in gonadotrophin release which occurs after ovariectomy is caused by steroid withdrawal resulting in an enhanced pituitary responsiveness to LH releasing hormone (LHRH) associated with increased LHRH release and pituitary LHRH binding. The effects of oestrogen replacement after ovariectomy and chronic treatment of intact rats with an oestrogen antagonist, tamoxifen, on LH release and in-vitro pituitary responses to LHRH have been investigated. Capsules containing crystalline oestradiol, implanted at the time of ovariectomy, completely inhibited the rise in LH release although pituitary responsiveness was greater after 10 days in the oestrogen-treated rats than in untreated ovariectomized controls. On day 4 after ovariectomy pituitary responses to LHRH were comparable in both treated and untreated groups although in both groups the responses were greater than those measured in intact dioestrous rats. Treatment with tamoxifen over a 4-day period also augmented pituitary responsiveness but only at the lowest dose (0·5 mg/kg); no effect on serum LH concentrations was observed. Higher doses of the antagonist (1 and 2 mg/kg) did not affect pituitary responses, although the highest dose did cause a significant rise in serum LH. Treatment with a daily dose of 50 ng [d-Ser(But)6]LHRH(1–9)nonapeptide-ethylamide, starting on the day of ovariectomy, markedly attenuated the LH responses to LHRH ex vivo at days 2, 4 and 10 after ovariectomy. In contrast, the analogue treatment did not abolish the rise in LH release but this was proportionately less than in controls.

Clomiphene citrate induces luteinizing hormone release through hypothalamic luteinizing hormone-releasing hormone in vitro

1983 ◽  
Vol 103 (3) ◽  
pp. 289-292 ◽  
Author(s):  
A. Miyake ◽  
K. Tasaka ◽  
T. Sakumoto ◽  
Y. Kawamura ◽  
Y. Nagahara ◽  
...  
Keyword(s):  
Luteinizing Hormone ◽  
Clomiphene Citrate ◽  
Female Rats ◽  
Normal Female ◽  
Releasing Hormone ◽  
Superfusion System ◽  
In Series ◽  
Lh Release ◽  
Lh Releasing Hormone

Abstract. The releasing effects of clomiphene citrate (clomiphene) on luteinizing hormone (LH) and LH-releasing hormone (LRH) were examined in a sequential double chamber superfusion system by superfusing the mediobasal hypothalami (MBH) and/or pituitaries excised from normal female rats in dioestrus. When the MBH and the pituitary were superfused in sequence with medium containing 2 × 10−10 m oestradiol (E2), two significant peaks in LH release (60–130% increase, P < 0.05) were observed 40 min and 90 min after the administration of 3 × 10−8 mol clomiphene. Administration of clomiphene in medium without E2 induced a low peak (25–50% increase, P < 0.05) of LH released from the pituitary perfused in series with the MBH. Administration of clomiphene did not cause a marked increase of LH from the pituitary superfused alone, when superfused with or without E2 containing medium. The concentration of LRH in the efflux was significantly increased (50–100%) 40 min and 90 min after clomiphene administration when MBH was superfused with medium containing E2, whereas clomiphene had no effect when superfused with medium alone. These data indicate: 1) that clomiphene induces LRH release from the MBH, that it may induce LH release, in part, by acting directly at the pituitary level; 2) that changes in LH after clomiphene administration coincide with LRH release, and 3) that a certain concentration of E2 may be necessary for the secretion of LRH by clomiphene.

Epidermal growth factor stimulates secretion of rat pituitary luteinizing hormone in vitro

1985 ◽  
Vol 108 (2) ◽  
pp. 175-178 ◽  
Author(s):  
Akira Miyake ◽  
Keiichi Tasaka ◽  
Shirou Otsuka ◽  
Hiroko Kohmura ◽  
Hiroshi Wakimoto ◽  
...  
Keyword(s):  
Growth Factor ◽  
Luteinizing Hormone ◽  
Epidermal Growth Factor ◽  
Female Rats ◽  
Gonadotrophin Secretion ◽  
Significant Release ◽  
Lh Release ◽  
Epidermal Growth ◽  
Lh Releasing Hormone

Abstract. The effects of epidermal growth factor (EGF) on pituitary luteinizing hormone (LH) release and on the releases induced by oestradiol (E2) and LH-releasing hormone (LRH) were examined in a sequential double chamber perifusion system. In this system the mediobasal hypothalami (MBH) and/or pituitaries excised from normally cycling female rats in dioestrus were perifused with test media. Perifusion with EGF at 1 ng/ml for 30 min induced significant release (80–100% increase, P <0.05) of LH from hypothalamo-pituitary pairs, but not from the pituitary alone. Perifusion of the pituitary alone with medium containing 1 ng/ml EGF, resulted in significant release of LH (70–140% increase, P < 0.05) after adminnistration of 10−7 m E2, but did not significantly influence LH release in response to 20 ng/ml LRH. These findings suggest that EGF may be involved in the regulation of pituitary gonadotrophin secretion by a direct effect on the hypothalamus and indirectly by increasing the pituitary responsiveness to E2.

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