Effect of pretreatment of long-term ovariectomized (OVX) rats with a competitive LRH antagonist on FSH release in vitro

1985 ◽  
Vol 109 (4) ◽  
pp. 481-484
Author(s):  
J. A. M. J. van Dieten ◽  
G. P. van Rees
Keyword(s):  
Specific Effect ◽  
High Concentration ◽  
Ovx Rats ◽  
High K ◽  
Basal Release ◽  
Pituitary Glands ◽  
Lh Release ◽  
Release In Vitro

Abstract. The effect of a single sc injection of an LRH antagonist ((Ac - D - p - Cl - Phe1,2,D-Trp3,D-Phe6,D-Ala10)-LRH, Org 30093) into OVX rats on FSH release 24 h later was studied. Plasma FSH was decreased but pituitary FSH content was not changed. Incubation of the pituitary glands during 4 h resulted in a decreased basal release. FSH release induced by a low concentration of LRH (1 ng/ml) was depressed but that of a high concentration (10000 ng/ml) was augmented in comparison to FSH release induced in control glands. However, pretreatment with the antagonist had no specific effect on FSH release in vitro induced by high K+ or high K+ plus mbcAMP and theophylline, indicating that the changes of pituitary responsiveness to LRH are not caused by those parts of the secretory mechanism which are stimulated by these secretagogues. Moreover, it is concluded that the changes of pituitary LH release induced by administration of an LRH antagonist also concern FSH.

Effect of pre-treatment of long-term ovariectomized rats with anti-LRH on the response of the pituitary gland in vitro to the analogue of LRH D-ser-(tBu)6-des-gly10-ethylamide-LRH (Buserelin®)

1983 ◽  
Vol 104 (3) ◽  
pp. 272-278 ◽  
Author(s):  
G. P. van Rees ◽  
J. A. M. J. van Dieten ◽  
J. de Koning ◽  
A. F. P. M. de Goey
Keyword(s):  
Pituitary Gland ◽  
Ovariectomized Rats ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Basal Release ◽  
Pituitary Glands ◽  
Lh Release ◽  
Pre Treatment

Abstract. Ovariectomized rats were injected iv with an antiserum against LRH or normal rabbit serum. AntiLRH caused a decrease of plasma LH and FSH. After 24 or 48 h, the rats were decapitated and the pituitary glands incubated in the presence of an analogue of LRH which reacts minimally with anti-LRH (Buserelin). Pretreatment with anti-LRH caused an increased response of pituitary LH release to Buserelin. Similar results were obtained with regard to FSH. In this case, however, basal release of FSH was lowered by pre-treatment with antiLRH. Pituitary LH and FSH contents were not affected by anti-LRH, but synthesis of LH and FSH in vitro was smaller than in control glands obtained from rats pretreated with normal rabbit serum.

Release and synthesis of FSH by pituitary glands of female rats in vitro

1982 ◽  
Vol 100 (4) ◽  
pp. 499-503 ◽  
Author(s):  
A. A. J. Jenner ◽  
J. de Koning ◽  
G. P. van Rees
Keyword(s):  
Protein Synthesis ◽  
Slow Rate ◽  
Female Rats ◽  
Intact Female ◽  
Ovx Rats ◽  
Basal Release ◽  
Pituitary Glands ◽  
Independent Synthesis ◽  
Intact Rats

Abstract. Anterior hemi-pituitary glands from intact female and ovariectomized (OVX) rats were incubated with or without a maximally effective dose of LRH. During an 8 h incubation, LRH-stimulated release of FSH by pituitary glands from intact rats was biphasic: an initial slow rate of release and, from 2 to 8 h, an enhanced rate of release. Basal release was low up to 4 h, after which a marked increase of the rate of release was measured: from 6 to 8 h there was no difference between the rates of basal and LRH-stimulated release. Basal and LRH-stimulated release of FSH by pituitary glands from OVX rats were high and approximately constant during an 8 h incubation. Both basal and LRH-stimulated release by glands from intact as well as OVX rats were protein synthesis dependent. During the incubations an LRH-independent synthesis of FSH was measured. The results suggest that this synthesis is involved, either directly or indirectly, in increasing the rate of basal release of FSH after 4 h. A comparison of release and synthesis of FSH with those of LH reveals characteristic differences.

Release of LH in vitro from anterior pituitary glands of ovariectomized rats by LRH or elevated potassium concentration after pre-treatment with oestradiol benzoate in vivo

1982 ◽  
Vol 99 (2) ◽  
pp. 206-210 ◽  
Author(s):  
A. M. I. Tijssen ◽  
J. de Koning ◽  
G. P. van Rees
Keyword(s):  
Ovariectomized Rats ◽  
Bicarbonate Buffer ◽  
High K ◽  
Oestradiol Benzoate ◽  
Negative Effect ◽  
Pituitary Glands ◽  
Lh Release ◽  
Pre Treatment ◽  
Positive Effect

Abstract. Pituitary glands from ovariectomized rats which had been pre-treated with oestradiol benzoate (OeB) or solvent oil were incubated in Krebs-Ringer bicarbonate buffer with glucose containing either LRH (1000 ng/ml) or a high K+ concentration (50 mM). OeB (7 μg sc) or oil was injected at 2.5 or 6.5 h before the beginning of the incubation experiment or during the three preceding days (three daily injections). Depending upon the period during which the pituitary glands had been exposed to OeB LH release induced by LRH was inhibited (negative effect of OeB) or augmented (positive effect). When the glands were incubated in medium containing high K+, only the negative effect of OeB pre-treatment was seen. It is concluded that that part of LRH-induced LH release which is mimicked by high K+ is involved in the negative effect of OeB, but not in its positive effect.

Effect of clomiphene citrate on the in vitro release of LH and FSH by the pituitary gland of the long-term ovariectomized rat pretreated with LRH or with LRH and oestradiol benzoate

1986 ◽  
Vol 113 (1) ◽  
pp. 35-41
Author(s):  
G. A. Schuiling ◽  
H. Moes ◽  
T. R. Koiter
Keyword(s):  
Clomiphene Citrate ◽  
Combined Treatment ◽  
In Vitro Release ◽  
Pituitary Gonadotropin ◽  
Ovx Rats ◽  
Oestradiol Benzoate ◽  
Pituitary Glands ◽  
Pre Treatment

Abstract. The effect of a combined in vivo pre-treatment with luteinizing hormone-releasing hormone (LRH) and either oestradiol benzoate (OB), clomiphene (-citrate) or OB plus clomiphene on the autonomous and the supramaximally LRH-stimulated in vitro secretion of LH and FSH by pituitary glands of long-term ovariectomized (OVX) rats was studied using a hemi-pituitary perifusion system. The concentration of LRH in the perifusion medium was 1 μg/ml. Pre-treatment with LRH during 5 days was effected by means of sc implanted Alzet® osmotic minipumps; control rats received a piece of silastic with the dimensions of a minipump. OB, 3 μg/injection, clomiphene 100 μg/injection or solvent were given on days 2 and 4 (day of perifusion: day 5). In rats not pre-treated with LRH neither OB, nor clomiphene changed the content of the pituitary gonadotropin stores. There was only a small but significant positive effect of the combined treatment with OB and clomiphene on the pituitary FSH content. LRH (partly) depleted the gonadotropin stores. This effect of LRH was potentiated by OB, but not by clomiphene. Clomiphene prevented the depletion-potentiating effect of OB. OB raised the LRH-stimulated secretion of LH and FSH as well as the autonomous secretion of LH. Clomiphene raised the LRH-stimulated (not the autonomous) secretion of LH and FSH. OB plus clomiphene had the same effect as OB alone. Clomiphene also raised the LRH-stimulated secretion of LH and FSH after pre-treatment with LRH, but OB did not do so: LRH prevented the stimulatory effect of OB but not of clomiphene. OB plus clomiphene had the same effect as OB alone. The absence of a stimulatory effect of OB on the LRH-stimulated secretion of LH and FSH in the LRH-pretreated rat appeared to be due to the very low gonadotropin content of the pituitary glands after pre-treatment with LRH and OB: the effect of OB on the LRH-responsiveness proper (i.e. release of LH and FSH as related to the pituitary LH and FSH content) remained stimulatory. Also clomiphene enhanced the LRH-responsiveness proper, but this drug cannot potentiate the gonadotropin stores-depleting effect of LRH. These results demonstrate that clomiphene exclusively 'behaves' like an oestrogen-agonist, able to enhance the LRH-stimulated gonadotropin secretion. Also in the LRH-pre-treated rat clomiphene acts like an oestrogen-agonist, but unlike oestradiol clomiphene cannot potentiate the LRH-induced depletion of the pituitary gonadotropin stores. Therefore, it can also raise the LRH-stimulated secretion of LH and FSH in the LRH-pre-treated OVX rat.

Effect of pretreatment of long-term ovariectomized rats with an LRH antagonist on LH release in vitro by LRH, elevated K+ or N6-monobutyryl adenosine 3',5'-monophosphate (mbcAMP) plus theophylline

1985 ◽  
Vol 108 (3) ◽  
pp. 331-337 ◽  
Author(s):  
G. P. van Rees ◽  
J. A. M.J. van Dieten
Keyword(s):  
Ovariectomized Rats ◽  
Serum Lh ◽  
Low Concentrations ◽  
Pituitary Glands ◽  
Lh Release ◽  
Release In Vitro ◽  
High Concentrations ◽  
Response Line ◽  
The Right

Abstract. The effect of the LRH antagonist (Ac-D-p--Cl-Phe1,2,D-Trp3,D-Phe6-D-Ala10)-LRH (Org 30093) on pituitary LH release was studied, using pituitary glands of ovariectomized rats. In vitro, the antagonist had no detectable agonist activity in the concentration used, had no effect on the maximal LH release which can be induced by LRH and shifted the dose-response line of LRH to the right, without changing its slope. By this the antagonist fulfilled the conditions of purely competitive antagonist. Also, when present in vitro, the antagonist had no effect on LH release induced by raised K+, whether alone or in combination with mbcAMP plus theophylline. A single injection of Org 30093 decreased serum LH without inducing a change of pituitary LH content measured 24 h later. Twenty-four h after the sc injection of the agonist, LH release in vitro induced by LRH was affected differently, depending on the concentration of LRH used: the effect of low concentrations of LRH was inhibited, whereas the effect of high concentrations of LRH was augmented. Pretreatment with the agonist had no effect on LH release by raised K+ combined with mbcAMP plus theophylline, but slightly increased LH release by raised K+.

The depressing (negative) effect of oestradiol benzoate on the in vitro secretion of LH and FSH by the pituitary gland of the LRH-pretreated long-term ovariectomized rat changes into the augmentative (positive) effect after discontinuation of the LRH-pretreatment

1985 ◽  
Vol 110 (3) ◽  
pp. 329-337 ◽  
Author(s):  
G. A. Schuiling ◽  
H. Moes ◽  
T. R. Koiter
Keyword(s):  
Depressing Effect ◽  
Ovx Rats ◽  
Gonadotrophin Secretion ◽  
Oestradiol Benzoate ◽  
Negative Effect ◽  
Positive Effect ◽  
Perifusion System

Abstract. The effect of pretreatment in vivo with oestradiol benzoate on in vitro secretion of LH and FSH was studied in long-term ovariectomized (OVX) rats both at the end of a 5-day continuous in vivo pretreatment with LRH and 4-days after cessation of such LRH pretreatment. Rats were on day 0 sc implanted with osmotic minipumps which released LRH at the rate of 250 ng/h. Control rats were implanted with a piece of silicone elastomer with the dimensions of a minipump. On days 2 and 4 the rats were injected with either 3 μg EB or with oil. On day 5 part of the rats were decapitated and the in vitro autonomous (i.e. non-LRH-stimulated) and 'supra-maximally' LRHstimulated release of LH and FSH was studied using a perifusion system. From other rats the minipumps were removed on day 5 and perifusion was performed on day 9. On the 5th day of the in vivo LRH pretreatment the pituitary LH/FSH stores were partially depleted; the pituitaries of the EB-treated rats more so than those of the oil-injected rats. EB alone had no significant effect on the content of the pituitary LH- and FSH stores. On day 9, i.e. 4 days after removal of the minipumps, the pituitary LH and FSH contents had increased in both the oil- and the EB injected rats, but had not yet recovered to control values. In rats not subjected to the 5-days pretreatment with LRH EB had a positive effect on the supra-maximally LRH-stimulated secretion of LH and FSH as well as on the non-stimulated secretion of LH. EB had no effect on the non-stimulated secretion of FSH. After 5 days of in vivo pretreatment with LRH only, the in vitro non-stimulated and supra-maximally LRH-stimulated secretion of both LH and FSH were strongly impaired, the effect correlating well with the LRH-induced depletion of the pituitary LH/FSH stores. In such LRH-pretreated rats EB had on day 5 a negative effect on the (already depressed) LRH-stimulated secretion of LH (not on that of FSH). EB had no effect on the non-stimulated LH/FSH secretion. It could be demonstrated that the negative effect of the combined LRH/EB pretreatment was mainly due to the depressing effect of this treatment on the pituitary LH and FSH stores: the effect of oestradiol on the pituitary LRH-responsiveness (release as related to pituitary gonadotrophin content) remained positive. In LRH-pretreated rats, however, this positive effect of EB was smaller than in rats not pretreated with LRH. Four days after removal of the minipumps there was again a positive effect of EB on the LRH-stimulated secretion of LH and FSH as well as on the non-stimulated secretion of LH. The positive effect of EB on the pituitary LRH-responsiveness was as strong as in rats which had not been exposed to exogenous LRH. The non-stimulated secretion of FSH was again not affected by EB. The results demonstrate that the effect of EB on the oestrogen-sensitive components of gonadotrophin secretion consists of two components: an effect on the pituitary LRH-responsiveness proper, and an effect on the pituitary LH/FSH stores. The magnitude of the effect of EB on the LRH-responsiveness is LRH dependent: it is very weak (almost zero) in LRH-pretreated rats, but strong in rats not exposed to LRH as well as in rats of which the LRH-pretreatment was stopped 4 days previously. Similarly, the effect of EB on the pituitary LH and FSH stores is LRH-dependent: in the absence of LRH, EB has no influence on the contents of these stores, but EB can potentiate the depleting effect of LRH on the LH/FSH-stores. Also this effect disappear after cessation of the LRH-pretreatment.

Effect of hypothalamic neuropeptides on corticotrophin release from quarters of rat anterior pituitary gland in vitro

1984 ◽  
Vol 100 (2) ◽  
pp. 219-226 ◽  
Author(s):  
S. A. Nicholson ◽  
T. E. Adrian ◽  
B. Gillham ◽  
M. T. Jones ◽  
S. R. Bloom
Keyword(s):  
Pituitary Gland ◽  
Anterior Pituitary ◽  
Low Dose ◽  
Physiological Role ◽  
Anterior Pituitary Gland ◽  
High Concentration ◽  
Response Curves ◽  
Corticotrophin Releasing Factor ◽  
Basal Release

ABSTRACT The effect of six hypothalamic peptides on the basal release of ACTH and that induced by arginine vasopressin (AVP) or by ovine corticotrophin releasing factor (oCRF) from fragments of the rat anterior pituitary gland incubated in vitro was investigated. Dose–response curves to AVP and to oCRF were obtained, and the response to a low dose of oCRF was potentiated by a low dose of AVP. Basal release of ACTH was not affected by any of the peptides in concentrations in the range 10−12 to 10−6 mol/l, and only substance P (SP) and somatostatin (SRIF) inhibited significantly the response to oCRF in a dose-related manner. The responses to a range of doses of oCRF or AVP were reduced by 10−8 and 10 − 6 mol SP or SRIF/1, and to a greater extent by the higher dose. Except in the case of 10−6 mol SRIF/1 on the response to AVP, the response was not further diminished by preincubation of the tissue with the peptide before the stimulating agent was added. The inhibition of the responses to AVP or oCRF by 10−9 mol SP/1 was not potentiated by its combination with either 5 × 10−10 or 10−8 mol SRIF/1; the inhibitory effects were merely additive. The results suggest that although SRIF and SP are able to modulate the release of ACTH from the anterior pituitary gland, they do so only at a high concentration. In the case of SRIF these concentrations are several orders of magnitude higher than those reported to be present in the hypophysial portal blood and therefore a physiological role for this peptide in the control of ACTH secretion is unlikely. J. Endocr. (1984) 100, 219–226

In-vitro control of growth hormone secretion by synthetic releasing factors in young and adult ducks (Anas platyrhynchos)

1988 ◽  
Vol 119 (3) ◽  
pp. 421-429 ◽  
Author(s):  
C. Foltzer-Jourdainne ◽  
S. Harvey ◽  
P. Mialhe
Keyword(s):  
Low Dose ◽  
Hormone Secretion ◽  
Growth Hormone Secretion ◽  
Age Related ◽  
Basal Release ◽  
Pituitary Glands ◽  
Releasing Factors ◽  
Somatostatin 14

ABSTRACT Release of GH from perifused duckling hemipituitaries was stimulated, in a biphasic manner, by synthetic TRH and human pancreatic GH-releasing factor (GRF). At all effective concentrations, the level of GH release was increased within 5 min of TRH or GRF perifusion and was maximal after 10 min of TRH perifusion and after 20 min of GRF perifusion. Although TRH was perifused for 20 min the level of GH release declined during the last 10 min. The most effective dose of TRH (1·0 μg/ml; 2·7 μmol/l) and GRF (0·5 μg/ml; 110 nmol/l) provoked similar (250– 300%) increases in the level of GH release. However, since the effect of TRH was only of short duration, the total release of GH induced by GRF was higher than that elicited by TRH, especially with the low dose. The increase in release of GH induced by TRH or GRF was blunted when pituitaries from adult ducks were used. As in young ducks, the GH response to GRF was higher, whereas the response to TRH was very low. The GH response of perifused adult pituitaries to GRF was, however, potentiated when TRH was perifused simultaneously. The basal release of GH from both young and adult pituitary glands was unaffected by perifusion with somatostatin-14 (SRIF-14) at doses of 1 and 2 μg/ml. The perifusion of hemipituitary glands with similar doses of SRIF-14 was also unable to suppress the stimulation of GH release induced by prior perifusion with GRF, although when SRIF-14 and TRH were simultaneously perifused TRH-induced GH release was markedly suppressed. These results demonstrate direct effects and interactions of TRH, GRF and SRIF on the release of GH from duck pituitary glands. GRF is the most potent releasing factor for GH in both young and adult ducks although in adult ducks it is less effective. These results also provide evidence that the age-related decline in the in-vivo GH response to TRH is due to a desensitization of pituitary somatotrophs. J. Endocr. (1988) 119, 421–429

LH-RH-dependent synthesis of protein necessary for LH release from rat pituitary glands in vitro

1976 ◽  
Vol 5 (1-2) ◽  
pp. 151-160 ◽  
Author(s):  
J. De Koning ◽  
J.A.M.J. Van Dieten ◽  
G.P. Van Rees
Keyword(s):  
Rat Pituitary ◽  
Pituitary Glands ◽  
Lh Release ◽  
Lh Rh
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