Prior exposure to gonadotrophins prevents the subsequent antigonadotrophic actions of cloprostenol by a cyclic AMP-dependent mechanism in cultured human granulosa cells

1991 ◽  
Vol 131 (2) ◽  
pp. 319-325 ◽  
Author(s):  
A. E. Michael ◽  
G. E. Webley
Keyword(s):  
Cyclic Amp ◽  
Corpus Luteum ◽  
Granulosa Cells ◽  
Early Pregnancy ◽  
Prior Exposure ◽  
Vitro Fertilization ◽  
Human Granulosa Cells ◽  
Dependent Mechanism

ABSTRACT The antigonadotrophic action of a prostaglandin F2α analogue, cloprostenol, has been investigated in human granulosa cells obtained from cycles stimulated for in-vitro fertilization and induced to secrete luteal quantities of progesterone by culture in serum-supplemented medium. Cells were exposed to conditions which may mimic those occurring in early pregnancy to establish the roles of human chorionic gonadotrophin (hCG) versus LH and that of cyclic AMP (cAMP) in the anti-gonadotrophic action of cloprostenol. When human granulosa cells were cultured in the absence of treatment for 3 days, exposure to cloprostenol had no effect on basal progesterone production but inhibited hCG-stimulated progesterone (60% decrease; P<0·01), hCG-stimulated cAMP (40% decrease; P < 0·05) and the progesterone response to dibutyryl cAMP (dbcAMP; 70% decrease; P < 0·01), suggesting pre- and post-cAMP sites of cloprostenol action. The inhibitory actions of cloprostenol were prevented when the granulosa cells were either continuously exposed to treatment from the start of culture or pre-exposed for 3 days to maximum concentrations of LH, hCG, dbcAMP or 8-bromo-cAMP. We conclude that prior exposure either in vivo or in vitro to LH or hCG prevents the subsequent antigonadotrophic action of cloprostenol via a cAMP-dependent mechanism. Prevention of the antigonadotrophic action of cloprostenol after exposure to hCG may be a mechanism through which CG prevents regression of the corpus luteum in early pregnancy, while the suppressive effect of LH pretreatment may account for the refractory response of the early corpus luteum to cloprostenol following the midcycle LH surge. Journal of Endocrinology (1991) 131, 319–325

In vitro Stimulation of Granulosa Cells by a Combination of Different Active Ingredients of Unkei-to

2004 ◽  
Vol 32 (04) ◽  
pp. 569-578 ◽  
Author(s):  
Wen-Shu Sun ◽  
Atsushi Imai ◽  
Keiko Tagami ◽  
Michiyo Sugiyama ◽  
Tatsuro Furui ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Response Curves ◽  
Vitro Fertilization ◽  
Human Granulosa Cells ◽  
Estradiol Secretion ◽  
Paeoniae Radix ◽  
In Vivo Effects ◽  
Stimulation Of

Unkei-to is widely used in traditional Japanese herbal medicine for its ovulation-inducing effect. In the present study, we investigated the in vivo effects of Unkei-to and its compounds on the steroidogenesis and cytokine secretion in human granulosa cells. Unkei-to stimulated the secretions of 17β-estradiol and progesterone from highly luteinized granulosa cells obtained from in vitro fertilization patients; the stimulated effect on estradiol secretion occurred with 0.3 μg/ml, while a significant effect on progesterone secretion was obtained at 10 μg/ml. The Unkei-to stimulation of estradiol secretion could be accounted for by the effects of its ingredients, Shakuyaku (paeoniae radix, Paeonia lactiflora Pallas) and Keihi (cinnamomi cortex, Cinnamomum cassia Blume); while dose response curves for Unkei-to and Keihi to induce progesterone production were superimposable. Exposure of the cells to Unkei-to caused dose-dependent increases in the concentrations of interleukin (IL)-1β, IL-6 and IL-8 in the culture medium. Similar results were obtained when cells were incubated with the ingredient Ninjin (ginseng radix, Panax ginseng C.A. Meyer), but not Shakuyaku and Keihi. These results indicate that Unkei-to has direct stimulatory effects on human granulosa cells to stimulate the steroidogenesis and secretion of cytokines (IL-1β, IL-6 and IL-8). The various beneficial actions of Unkei-to on the ovary may result from a combination of different ingredient herbs with different stimulatory effects on both steroidogenesis and the ovulatory process within the ovary, as well as stimulatory effect on the hypothalamus-pituitary axis.

Effect of gonadotrophins on progesterone secretion by cultured granulosa cells obtained from human preovulatory follicles

1985 ◽  
Vol 110 (3) ◽  
pp. 401-407 ◽  
Author(s):  
T. Hillensjö ◽  
A. Sjögren ◽  
B. Strander ◽  
L. Nilsson ◽  
M. Wikland ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Maximal Effect ◽  
Tubal Infertility ◽  
Vitro Fertilization ◽  
Follicular Maturation ◽  
Progesterone Secretion ◽  
Preovulatory Follicles ◽  
Ultrasound Guided Puncture

Abstract. Granulosa cells were obtained from human preovulatory follicles in 31 women undergoing in vitro fertilization and embryo transfer due to tubal infertility. Follicular maturation was stimulated and synchronized by treatment with Clomiphene or human menopausal gonadotrophin (hMG), or both, plus human chorionic gonadotrophin (hCG). Follicles were aspirated by ultrasound guided puncture approximately 34–36 h after the hCG injection. The granulosa cells were washed and suspended in modified medium 199 containing 10% foetal bovine serum and cultured as monolayers for 6–8 days in the absence and presence of hormones and reactants. Progesterone formation was analyzed by RIA. In general, the cells underwent morphological luteinization and secreted high amount of progesterone. Under basal conditions the secretion of progesterone was highest during the first 2 days in culture and then gradually declined. Progesterone secretion was stimulated by human LH, hCG and the adenylate cyclase stimulator forskolin, with a maximal effect between days 2–6. The β-adrenergic agonist isoproteronol in preliminary experiments potentiated the stimulatory effect of hCG but had no own stimulatory effect. No clear differences in progesterone secretion or responsiveness to in vitro stimulation relating to the various in vivo stimulation protocols were found.

scholarly journals Stimulation of Apoptosis in Human Granulosa Cells from in Vitro Fertilization Patients and Its Prevention by Dexamethasone: Involvement of Cell Contact and Bcl-2 Expression

2002 ◽  
Vol 87 (7) ◽  
pp. 3441-3451 ◽  
Author(s):  
Ravid Sasson ◽  
Abraham Amsterdam
Keyword(s):  
In Vitro Fertilization ◽  
Actin Cytoskeleton ◽  
Gap Junctions ◽  
Granulosa Cells ◽  
Connexin 43 ◽  
Pronounced Increase ◽  
Vitro Fertilization ◽  
Human Granulosa Cells ◽  
Progesterone Production

Human granulosa cells obtained from in vitro fertilization patients are highly luteinized, but can still be stimulated by LH/cAMP for production of progesterone. This stimulation involved enhancement of apoptosis. Incubation of the cells with dexamethasone (Dex) reduced the apoptotic incidence compared with nontreated cells and completely abolished the increase in apoptosis stimulated by LH or forskolin, concomitantly with a pronounced increase in progesterone production. Organization of the actin cytoskeleton was dramatically reduced after LH/forskolin stimulation. In contrast, Dex prevented disorganization of the actin filament networks. LH and forskolin also decreased the organization of gap junctions, which could be prevented by Dex. However, the intracellular level of connexin 43 was elevated in the presence of LH, forskolin, and Dex. Endogenous levels of the survival gene protein Bcl-2 were significantly elevated in all cultures treated with Dex compared with either nonstimulated cultures or cultures stimulated with LH and forskolin. Our data suggest that LH/cAMP can stimulate steroidogenesis even during the initial stage of apoptosis of human granulosa cells, whereas Dex, which blocks apoptosis, could further elevate progesterone production. Moreover, the integrity of gap junctions and the actin cytoskeleton as well as elevated levels of Bcl-2 may play an important role in the suppression of apoptosis of human granulosa cells.

Effect of noradrenaline and dopamine on progesterone and estradiol secretion of human granulosa cells

1993 ◽  
Vol 129 (2) ◽  
pp. 165-168 ◽  
Author(s):  
József Bódis ◽  
Hans R Tinneberg ◽  
Attila Török ◽  
Philippe Cledon ◽  
Volker Hanf ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Direct Action ◽  
Serum Free Medium ◽  
Free Medium ◽  
Vitro Fertilization ◽  
Serum Free ◽  
Human Granulosa Cells ◽  
Estradiol Secretion ◽  
Cells Cultured

The aim of this study was to explore the direct action of noradrenaline and dopamine on progesterone and estradiol secretion of human granulosa cells cultured in serum-free medium. Progesterone and estradiol production was measured in the presence and absence of noradrenaline, dopamine or propranolol using radioimmunoassays; statistical analysis was performed by analysis of variance and Newman-Keul's multiple range test. Twenty-six women aged 31±3 years undergoing in vitro fertilization and embryo transfer for infertility treatment at University Women's Hospital, University of Tübingen, Germany, took part in this study. Noradrenaline significantly inhibited progesterone production by human granulosa cells in a dose-related manner at a concentration of 10−4–10−6 mol/l. Dopamine significantly stimulated estradiol secretion by granulosa cells in an inverse dose-related manner. Both effects were blocked by propranolol. The results suggest that catecholaminergic actions switch over the steroid production of human granulosa cells cultured in serum-free medium from progesterone to estradiol.

In vivo and in vitro effect of growth hormone on estradiol secretion by human granulosa cells.

1993 ◽  
Vol 77 (1) ◽  
pp. 61-67 ◽  
Author(s):  
A Barreca ◽  
P G Artini ◽  
P Del Monte ◽  
P Ponzani ◽  
P Pasquini ◽  
...  
Keyword(s):  
Growth Hormone ◽  
Granulosa Cells ◽  
Human Granulosa Cells ◽  
Estradiol Secretion ◽  
Vitro Effect
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