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

2019 ◽  
Vol 54 (9) ◽  
pp. 1236-1243 ◽  
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

Formation of 4-oestrene-3,17-dione (19-norandrostenedione) by porcine granulosa cells in vitro is inhibited by the aromatase inhibitor 4-hydroxyandrostenedione and the cytochrome P-450 inhibitors aminoglutethimide phosphate and ketoconazole

1989 ◽  
Vol 120 (2) ◽  
pp. 251-260 ◽  
Author(s):  
M. W. Khalil ◽  
P. Morley ◽  
M. A. Glasier ◽  
D. T. Armstrong ◽  
T. Lang
Keyword(s):  
High Performance Liquid Chromatography ◽  
Aromatase Inhibitor ◽  
Granulosa Cells ◽  
High Performance ◽  
Reverse Phase ◽  
Porcine Granulosa Cells ◽  
Cytochrome P 450 ◽  
Ovarian Follicular Fluid ◽  
Cells Cultured

ABSTRACT The origin and biosynthesis of 4-oestrene-3,17-dione (19-norandrostenedione), a major steroid in porcine ovarian follicular fluid, was investigated by culturing granulosa cells from 4–6 mm follicles of prepubertal gilts with radiolabelled androstenedione and 19-hydroxyandrostenedione. Steroid metabolites were purified by solvent extraction and lipophilic column chromatography, and analysed by C18 reverse-phase high-performance liquid chromatography. 19-Hydroxyandrostenedione, 19-norandrostenedione and oestradiol-17β were obtained as major metabolites from androstenedione, while 19-norandrostenedione and oestradiol-17β were the major products from 19-hydroxyandrostenedione. Serum alone or serum plus FSH significantly enhanced formation of 19-norandrostenedione and oestradiol-17β from each substrate, compared with controls. Micromolar concentrations (1 μmol/l) of 4-hydroxyandrostenedione, an aromatase inhibitor, significantly reduced formation of 19-norandrostenedione and oestradiol-17β by granulosa cells cultured with serum and FSH. Formation of 19-norandrostenedione and oestradiol-17β from androstenedione and 19-hydroxyandrostenedione was also significantly inhibited by aminoglutethimide phosphate, a cytochrome P-450 inhibitor known to block the conversion of androstenedione to oestrogens. Ketoconazole, an inhibitor of the cytochrome P-450 dependent 17,20-lysase, blocked formation of 19-norandrostenedione and oestradiol-17β only at millimolar concentrations. These results suggest that 19-norsteroid and oestrogen formation from C19 aromatizable androgens may share a common or overlapping pathway, and imply that 19-norsteroid and oestrogen synthesis is mediated by cytochrome P-450 dependent enzymes. Journal of Endocrinology (1989) 120, 251–260

Porcine granulosa cell conditioned media as autocrine regulator of progesterone secretion

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.

Gonadotrophic regulation of prolactin mediated progesterone secretion in vitro

1985 ◽  
Vol 110 (2) ◽  
pp. 251-256 ◽  
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.

scholarly journals Phospholipase C inhibits apoptosis of porcine primary granulosa cells cultured in vitro

2019 ◽  
Vol 12 (1) ◽  
Author(s):  
Huali Chen ◽  
Youfu Yang ◽  
Youlin Wang ◽  
Yuan Li ◽  
Yamei He ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Phospholipase C ◽  
Granulosa Cells ◽  
Protein Abundance ◽  
Kinase C ◽  
Early Apoptosis ◽  
Dependent Protein ◽  
Late Apoptosis ◽  
Cells Cultured

Abstract Phospholipase C (PLC) can participate in cell proliferation, differentiation and aging. However, whether it has a function in apoptosis in porcine primary granulosa cells is largely uncertain. The objective of this study was to examine the effects of PLC on apoptosis of porcine primary granulosa cells cultured in vitro. The mRNA expression of BAK, BAX and CASP3, were upregulated in the cells treated with U73122 (the PLC inhibitor). The abundance of BCL2 mRNA, was upregulated, while BAX and CASP3 mRNA expression was decreased after treatment with m-3M3FBS (the PLC activator). Both the early and late apoptosis rate were maximized with 0.5 μM U73122 for 4 h. The rate of early apoptosis was the highest at 4 h and the rate of late apoptosis was the highest at 12 h in the m-3M3FBS group. The protein abundance of PLCβ1, protein kinase C β (PKCβ), calmodulin-dependent protein kinaseII α (CAMKIIα) and calcineurinA (CalnA) were decreased by U73122, and CAMKIIα protein abundance was increased by m-3M3FBS. The mRNA expression of several downstream genes (CDC42, NFATc1, and NFκB) was upregulated by PLC. Our results demonstrated that apoptosis can be inhibited by altering PLC signaling in porcine primary granulosa cells cultured in vitro, and several calcium−sensitive targets and several downstream genes might take part in the processes.

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

2021 ◽  
pp. 1-9
Author(s):  
Christina I. Messini ◽  
Anna Vasilaki ◽  
Evangelia Korona ◽  
George Anifandis ◽  
Eleni Katsiani ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Progesterone Secretion
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