Effect of adiponectin on estradiol and progesterone secretion from human luteinized granulosa cells in vitro

Daidzein, a phytoestrogen present in soybean products used in swine feed, has been demonstrated to affect both reproductive and endocrine functions. The aims of this study were to examine the in vitro effects of daidzein on (1) progesterone (P4) and oestradiol (E2) secretion by porcine luteinised granulosa cells harvested from medium follicles, and (2) the mRNA and protein expression of oestrogen receptors α and β (ERα and ERβ) in these cells. The influence of E2 on P4 secretion and ERα and ERβ expression in the granulosa cells of pigs was also investigated. It was found that daidzein inhibited progesterone secretion by luteinised granulosa cells isolated from medium follicles. In contrast, E2 did not affect progesterone production by these cells. Moreover, daidzein did not alter the granulosal secretion of E2. Both daidzein and E2 decreased mRNA expression of ERα in the cells examined. The expression of ERβ mRNA was not affected by daidzein but was inhibited by E2. ERα protein was not detected while ERβ protein was found in the nuclei of the cells. Daidzein and E2 upregulated the expression of ERβ protein in the cells. In summary, the phytoestrogen daidzein directly affected the porcine ovary by inhibiting progesterone production and increasing ERβ protein expression. Daidzein-induced changes in follicular steroidogenesis and granulosal sensitivity to oestrogens may disturb reproductive processes in pigs.

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

Acta Endocrinologica ◽

10.1530/acta.0.1100401 ◽

1985 ◽

Vol 110 (3) ◽

pp. 401-407 ◽

Cited By ~ 13

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.

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Regulation and action of fibroblast growth factor 17 in bovine follicles

Journal of Endocrinology ◽

10.1677/joe-09-0145 ◽

2009 ◽

Vol 202 (3) ◽

pp. 347-353 ◽

Cited By ~ 39

Author(s):

M F Machado ◽

V M Portela ◽

C A Price ◽

I B Costa ◽

P Ripamonte ◽

...

Keyword(s):

Growth Factor ◽

Fibroblast Growth Factor ◽

Granulosa Cells ◽

Cumulus Cells ◽

Mrna Abundance ◽

Fibroblast Growth ◽

Theca Cells ◽

Progesterone Secretion ◽

Igf I

Fibroblast growth factor 17 (FGF17) is a member of the FGF8 subfamily that appears to be relevant to folliculogenesis and oogenesis, as the prototype member FGF8 is an oocyte-derived protein that signals to cumulus cells. FGF8 has structural and receptor-binding similarities to FGF17, whose expression in the ovary has not been reported. In this study, we demonstrate localization of FGF17 protein to the oocyte of preantral follicles, and to the oocyte and granulosa cells of antral follicles. Real-time PCR demonstrated the presence of mRNA in oocytes and, to a lesser extent, in granulosa and theca cells. FGF17 mRNA abundance was low in granulosa and theca cells from healthy follicles and increased significantly in atretic follicles. Addition of FSH or IGF-I to granulosa cells in vitro decreased FGF17 mRNA abundance, and treatment with FGF17 inhibited estradiol and progesterone secretion from granulosa cells in relation to control cultures without these additives. We conclude that FGF17 is a potential mediator of granulosa cell differentiation.

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Granulosa cells of the cumulus oophorus are different from mural granulosa cells in their response to gonadotrophins and IGF-I

Journal of Endocrinology ◽

10.1677/joe.0.1700565 ◽

2001 ◽

Vol 170 (3) ◽

pp. 565-573 ◽

Cited By ~ 23

Author(s):

F Khamsi ◽

S Roberge

Keyword(s):

Granulosa Cells ◽

Cumulus Cells ◽

Saline Control ◽

Cell Replication ◽

Progesterone Secretion ◽

Igf I ◽

Negative Effect ◽

Positive Effect

There are two types of granulosa cells: those which surround the oocyte are cumulus cells (CC) and those which surround the antrum are mural granulosa cells (MGC). These cells are under the influence of several hormones and growth factors, the most important of which are gonadotrophins and IGF-I. In this article, we report novel observations on the differences between these two types of granulosa cells and their interaction with gonadotrophins and IGF-I. We were able to conduct physiological studies on the role of IGF-I by using an analogue of IGF-I which does not bind to IGF-I-binding proteins (LR3-IGF-I). Immature rats received saline, equine chorionic gonadotrophin (eCG), LR3-IGF-I or eCG plus LR3-IGF-I by infusion using a pump from 24-29 days of age. The rats were killed and the ovaries removed. Surface follicles were punctured and MGC and oocyte cumulus complexes were removed. These were cultured in saline (control) and in three different doses of FSH. Cell replication was assessed by 3H-thymidine incorporation and differentiation was evaluated by the measurement of progesterone secretion. It was noted that CC replicated ten times more than MGC. Similarly, progesterone secretion by CC was six times more than by MGC. In vivo exposure to gonadotrophins (eCG) positively influenced in vitro treatment with FSH in both cell types. This phenomenon was observed in both cell replication and progesterone secretion. The IGF-I analogue had a positive effect on cell replication of MGC but a negative effect on the cell replication of CC. With respect to progesterone secretion, the IGF-I analogue had a negative effect on CC but a positive effect on MGC. In conclusion, CC behaved differently from MGC in response to gonadotrophins and the IGF-I analogue. IGF-I and FSH acted additively, synergistically or antagonistically in different circumstances.

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Porcine granulosa cell conditioned media as autocrine regulator of progesterone secretion

Reproduction Fertility and Development ◽

10.1071/rd9930095 ◽

1993 ◽

Vol 5 (1) ◽

pp. 95

Author(s):

RT Denkova ◽

IG Ivanov ◽

LN Kanchev

Keyword(s):

Granulosa Cell ◽

Granulosa Cells ◽

Cultured Cells ◽

Primary Cultures ◽

Inhibitory Effect ◽

Conditioned Media ◽

Progesterone Secretion ◽

Porcine Granulosa Cells ◽

Porcine Granulosa Cell

The ability of porcine granulosa cells to release a progesterone inhibiting substance(s) was examined in vitro. Granulosa cells (SGCs, MGCs and LGCs) were harvested from small, medium or large antral follicles respectively. The effect of granulosa cell conditioned media obtained from small follicles (SGCCM) was studied in the culture of LGCs by estimation of progesterone secretion; the conditioned media evoked the inhibition of progesterone secretion by the LGCs. SGCCM produced by various numbers of cultured granulosa cells showed a dose-related inhibition of progesterone production. A maximum inhibitory effect was noted when a 5-fold concentration of SGCCM was used. The addition of SGCCM had no effect on the growth of the cultured cells. The factor(s) inhibiting progesterone secretion appeared to be a nonsteroidal substance of molecular mass greater than 10 kDa and was heat-stable and trypsin-sensitive. The data presented support the suggestion that the conditioned media generated by primary cultures of SGCs contain nonsteroidal regulators capable of inhibiting progesterone secretion by cultured LGCs; this inhibitory activity can play an important autocrine regulatory role in the process of follicular differentiation.

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Gonadotrophic regulation of prolactin mediated progesterone secretion in vitro

Acta Endocrinologica ◽

10.1530/acta.0.1100251 ◽

1985 ◽

Vol 110 (2) ◽

pp. 251-256 ◽

Cited By ~ 3

Author(s):

J. Steven Alexander ◽

Thomas M. Crisp

Keyword(s):

Granulosa Cells ◽

In Vitro Model ◽

Fold Increase ◽

Model System ◽

Regulatory Mechanisms ◽

Progesterone Secretion ◽

Vitro Model ◽

Dose Dependent ◽

Cells Cultured

Abstract. The effects of preincubating rat granulosa cells with FSH, LH, and Prl on subsequent Prl mediated progesterone secretion were investigated. Granulosa cells were isolated from ovarian follicles 50 h after injection of 5 IU PMSG and were then plated on poly-l-lysine coated coverslips in serum supplemented medium. Cells were preincubated for 24 h in the absence of hormones (control) or with the addition of either 0.25, 2.5, 25 ng/ml rat FSH or rat LH, or 1 μg/ml rat Prl. Following the preincubation period, cells were maintained for an additional 6 or 8 days in the presence or absence of 1 μg/ml Prl. When cells were preincubated with FSH or LH, only the two higher concentrations (2.5 and 25 ng/ml) stimulated significantly more progesterone secretion than control cultures during the 24 h preincubation period. For each series of preincubations, cells cultured for 6 or 8 days in the presence of Prl secreted significantly more progesterone at each day of culture than cells cultured without Prl. Cells preincubated and cultured with Prl secreted only 3–7-fold more progesterone than cells preincubated in control medium and then cultured with Prl. Preincubation with FSH or LH promoted a 20–45-fold increase in Prl mediated progesterone secretion compared to control preincubation cultures that also subsequently were cultured with Prl. The magnitude of Prl mediated progesterone secretion observed through 6 days of culturing was dose dependent on the preincubation concentration of FSH or LH. The establishment of an in vitro model system in which gonadotrophins enhance the responsiveness of granulosa cells to Prl in serum supplemented medium provides the opportunity for study of the regulatory mechanisms involved with the induction and maintenance of such responsiveness.

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The effects of phospholipase C on oestradiol and progesterone secretion in porcine granulosa cells cultured in vitro

Reproduction in Domestic Animals ◽

10.1111/rda.13517 ◽

2019 ◽

Vol 54 (9) ◽

pp. 1236-1243 ◽

Cited By ~ 1

Author(s):

Huali Chen ◽

Youfu Yang ◽

Youlin Wang ◽

Yamei He ◽

Jiaxin Duan ◽

...

Keyword(s):

Phospholipase C ◽

Granulosa Cells ◽

Progesterone Secretion ◽

Porcine Granulosa Cells ◽

Cells Cultured

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EFFECT OF LUTEINIZING HORMONE ON PROGESTERONE SECRETION IN VITRO BY THE GRANULOSA CELLS OF THE DOMESTIC FOWL (GALLUS DOMESTICUS)

Journal of Endocrinology ◽

10.1677/joe.0.0840249 ◽

1980 ◽

Vol 84 (2) ◽

pp. 249-254 ◽

Cited By ~ 28

Author(s):

J. W. WELLS ◽

A. B. GILBERT ◽

J. CULBERT

Keyword(s):

Luteinizing Hormone ◽

Granulosa Cells ◽

Peripheral Blood ◽

Domestic Fowl ◽

Gallus Domesticus ◽

Preovulatory Follicle ◽

Progesterone Secretion ◽

Physicochemical Methods

During short periods of incubation (3 h) the secretion of progesterone by granulosa cells from the largest preovulatory follicle of the fowl was higher (160 pmol/μg DNA) with ovine LH in the medium than without it (60 pmol/μg DNA). Granulosa cells from follicles collected 24 and 48 h before their expected ovulation secreted progesterone at similar rates to cells from the largest follicle which was likely to ovulate within 5 h. The identity of progesterone was confirmed by physicochemical methods. After granulosa cells had been incubated with LH in Medium 199 for 24 h, the concentration of progesterone in the medium was 1·65 μmol/l whereas oestrone and oestradiol were present at concentrations of 254 and 199 pmol/l respectively. The results indicate that the larger yellow yolk-filled follicles of the ovarian hierarchy in the domestic fowl contribute to the preovulatory surge of progesterone which has been observed in the peripheral blood.

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Bisphenol S Impaired Human Granulosa Cell Steroidogenesis in Vitro

International Journal of Molecular Sciences ◽

10.3390/ijms21051821 ◽

2020 ◽

Vol 21 (5) ◽

pp. 1821 ◽

Cited By ~ 1

Author(s):

Sarah Amar ◽

Aurélien Binet ◽

Ophélie Téteau ◽

Alice Desmarchais ◽

Pascal Papillier ◽

...

Keyword(s):

Granulosa Cells ◽

Messenger Rna ◽

Cellular Proliferation ◽

Structural Analog ◽

Bisphenol S ◽

Vitro Fertilization ◽

Progesterone Secretion ◽

Nanomolar Range ◽

Follicular Fluids

Bisphenol S (BPS) is a structural analog of the endocrine disruptor bisphenol A (BPA); it is the main BPA replacement in the plastics industry. Previous studies have shown that BPA and BPS exhibit similar effects on reproduction in fish and rodent species. BPS reportedly alters steroidogenesis in bovine granulosa cells. Luteinised granulosa cells collected from 59 women who were undergoing an in vitro fertilization procedure were cultured for 48 h in the presence or absence of BPS (10 nM, 100 nM, 1 µM, 10 µM or 50 µM). BPS exposure was investigated by assessing follicular fluids from these 59 women for their BPS content. Culture medium, cells, total messenger RNA (mRNA) and total protein extracted from the luteinised granulosa cells were examined for oestradiol and progesterone secretion, cellular proliferation, viability, gene expression, steroidogenic enzyme expression and cell signaling. BPS was measured in follicular fluids using mass spectrometry. Exposure of granulosa cells to 10 or 50 µM BPS for 48 h induced a 16% (p = 0.0059) and 64% (p < 0.0001) decrease, respectively, in progesterone secretion; 50 µM BPS decreased oestradiol secretion by 46% (p < 0.0001). Ten µM BPS also tended to reduce CYP11A1 protein expression by 37% (p = 0.0947) without affecting HSD3B1 and CYP19A1 expression. Fifty µM BPS increased ERRγ expression. Environmental levels of BPS (nanomolar range) did not induce changes in steroidogenesis in human granulosa cells. The effects of BPS were observed after only 48 h of BPS exposure. These acute effects might be similar to chronic effects of physiological BPS levels.

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The effect of human chorionic gonadotrophin on the expression of progesterone receptors in human luteal cells in vivo and in vitro

Reproduction ◽

10.1530/rep.1.00216 ◽

2005 ◽

Vol 130 (1) ◽

pp. 83-93 ◽

Cited By ~ 20

Author(s):

W Colin Duncan ◽

Eva Gay ◽

Jacqueline A Maybin

Keyword(s):

Granulosa Cells ◽

Luteal Phase ◽

Progesterone Receptors ◽

Chorionic Gonadotrophin ◽

Human Chorionic Gonadotrophin ◽

Progesterone Secretion ◽

Late Luteal Phase ◽

Lutein Cell

The human corpus luteum expresses genomic progesterone receptors (PRs) suggesting that progesterone may have an autocrine or paracrine role in luteal function. We hypothesised that the reduction in luteal PR reported in the late-luteal phase augmented progesterone withdrawal and had a role in luteolysis. We therefore tested the hypothesis that luteal rescue with human chorionic gonadotrophin (hCG) would maintain PR expression. PR was immunolocalised to different cell types in human corpora lutea (n = 35) from different stages of the luteal phase and after luteal rescue with exogenous hCG. There was no change in the staining intensity of theca-lutein cell or stromal cell PR throughout the luteal phase or after luteal rescue. In the late-luteal phase, granulosa-lutein cell PR immunostaining was reduced (P < 0.05) but the trend to reduction was also seen after luteal rescue with hCG (P = 0.055). To further investigate the effect of hCG on granulosa-lutein cell PR expression, an in vitro model system of cultured human luteinised granulosa cells was studied. Cells were cultured for 12–13 days exposed to different patterns of hCG and aminoglutethamide to manipulate progesterone secretion (P < 0.0001). Expression of PR A/B and PR B isoforms was examined by quantitative real-time RT-PCR. PR A/B mRNA was lower (P < 0.05) after 11–13 days of culture than after 7 days of culture. This reduction could not be prevented by hCG in the presence (P < 0.05) or absence (P < 0.05) of stimulated progesterone secretion. The expression of PR B mRNA showed a similar pattern (P = 0.054). Simulated early pregnancy in vivo and hCG treatment of luteinised granulosa cells in vitro did not appear to prevent the down-regulation of PR seen during luteolysis.

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