Follicle-stimulating hormone-induced estradiol and progesterone production by bovine antral and mural granulosa cells cultured in vitro in a completely defined medium.

Rouillier, P.; Matton, P.; Sirard, Marc‐André; Guilbault, L.A.

doi:10.2527/1996.74123012x

Cited by 30 publications

(47 citation statements)

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“…This is consistent with the response of these cells with respect to oestradiol secretion (Rouillier et al 1996). These data add to the literature by showing that protein as well as steroid hormones are affected by location of cells within the follicle (Roberts & Echternkamp 1994, Rouillier et al 1996.…”

Section: Discussionsupporting

confidence: 79%

“…Granulosa cells were harvested from 3-to 4-month-old Holstein prepubertal female calves as described (Rouillier et al 1996); in this culture system, aromatase activity decreases with time of culture but cells remain responsive to FSH. Animals were injected with 1500 IU equine chorionic gonadotrophin (eCG) (Equinex; Ayrest, Montreal, Canada) and ovaries were recovered by ovariectomy 96 h later.…”

Section: Methodsmentioning

confidence: 99%

“…Animals were injected with 1500 IU equine chorionic gonadotrophin (eCG) (Equinex; Ayrest, Montreal, Canada) and ovaries were recovered by ovariectomy 96 h later. An antral granulosa cell suspension was prepared according to the method described by Saumande (1991) and modified by Rouillier et al (1996); follicles d8 mm in diameter were punctured with an 18-gauge needle and rinsed ten times with 0·5 ml Menezo B 2 medium supplemented with heparin (100 IU/ml; Organon Teknika, Toronto, Ontario, Canada). Following removal of the antral cell suspension, the collapsed follicles were dissected and the internal wall was scraped gently with a smooth inoculation loop to remove the adherent mural granulosa cells.…”

Section: Methodsmentioning

confidence: 99%

“…Oestradiol and progesterone concentrations measured in conditioned medium from Experiment 1 have been reported for treatments with 0, 2 and 10 ng/ml FSH (Rouillier et al 1996). Oestradiol (Price et al 1995a) and progesterone (Lafrance & Goff 1985) were measured in medium samples in duplicate from day 6 of Experiment 2 without extraction as described.…”

Section: Assaysmentioning

confidence: 99%

“…We tested the hypothesis that FSH stimulates dimeric inhibin-A secretion from nonluteinized bovine granulosa cells, using two different culture systems in which oestradiol secretion is responsive to FSH. A further objective was to determine if subpopulations of granulosa cells (antral vs mural) differ in their ability to secrete dimeric inhibin-A, as they differ in their ability to secrete oestradiol (Rouillier et al 1996(Rouillier et al , 1998.…”

Section: Introductionmentioning

confidence: 99%

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Effect of FSH and cell localization on dimeric inhibin-A secretion from bovine granulosa cells in culture

Boudjemaa

Rouillier²,

Bhatia³

et al. 2000

Journal of Endocrinology

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We tested the hypotheses that the secretion of dimeric inhibin-A from cultured bovine granulosa cells is stimulated by FSH, and that antral cells secrete more inhibin-A than do mural cells. Cells from the antral or mural compartment of follicles were cultured in defined medium in two culture systems, and dimeric inhibin-A was measured by two-site ELISA or by Western immunoblotting. In the first culture system, dimeric inhibin-A secretion declined with time in culture, but was significantly (P<0·05) higher from antral than from mural cells (as was total inhibin-measured by RIA). The secretion of dimeric inhibin-A and inhibin-from antral but not mural cells was responsive to FSH. In the second culture system, dimeric inhibin-A secretion increased with time in culture, and was significantly stimulated by FSH, but FSH responsiveness was dependent on the concentrations of insulin in the culture medium. The major forms of inhibin-A secreted had molecular masses of approximately 58, 62, 103-116 and >116 kDa; the 32 kDa form was barely detectable. These different forms were all stimulated by FSH, but the >116 and 62 kDa forms were most responsive to FSH. We conclude that (i) FSH stimulates dimeric inhibin-A secretion from bovine granulosa cells, (ii) the 62 kDa form of inhibin-A may be more responsive to FSH than the 58 kDa form, and (iii) the spatial differentiation of granulosa cell function within the follicle previously observed for oestradiol secretion was also observed for inhibin-and dimeric inhibin-A secretion.

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Section: Discussionsupporting

confidence: 79%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Assaysmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Effect of FSH and cell localization on dimeric inhibin-A secretion from bovine granulosa cells in culture

Boudjemaa

Rouillier²,

Bhatia³

et al. 2000

Journal of Endocrinology

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Steroid production, cell proliferation, and apoptosis in cultured bovine antral and mural granulosa cells: Development of an in vitro model to study estradiol production

et al. 1998

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This study was undertaken to characterize the relationship between changes in steroid production, cell cycle activity (ie, cell proliferation) and apoptosis in antral and mural bovine granulosa cells cultured in vitro. This was done to select conditions promoting optimal estradiol production by bovine granulosa cells cultured in completely defined conditions. In the first experiment, antral granulosa cells were cultured over the entire 4 days of the culture period in the presence of either 0, 2, or 10 ng/ml of FSH (chronic conditions) or were maintained under minimal FSH support (0.5 ng/ml FSH) for the first 3 days of culture and then were challenged over the fourth day of culture with either 0, 2, or 10 ng/ml FSH (challenged conditions). Compared with cells exposed to constant FSH levels (chronic conditions), the FSH‐induced production of estradiol was higher (P < 0.006) and that of progesterone was lower (P < 0.02) over the last 24 h of culture, when antral granulosa cells were maintained under minimal FSH support during the first 3 days of culture (challenged conditions). In the second experiment, dynamics of estradiol and progesterone productions, conversion of [14C]androstenedione into subsequent steroid metabolites, DNA content, cell cycle activity, and apoptosis (as assessed by flow cytometry) of antral and mural granulosa cells over the first 3 days of culture under minimal FSH support and in response to a challenge with FSH during the last 24 h of culture were evaluated. Estradiol production as well as the conversion of androstenedione into testosterone and estradiol were greater (P< 0.01) in antral than in mural granulosa cells cultured under challenged conditions. A higher proportion of mural than antral granulosa cells were in the proliferative state at the end of culture (P < 0.03). This may be related to the decreased ability of mural cells to produce estradiol. FSH suppressed (P < 0.05) the spontaneous onset of apoptosis in both cell types. These results suggest that functional differences between these two cell compartments need to be considered in studying bovine granulosa cells in vitro. Because of their large (400 to 600%) FSH‐induced estradiol production, antral granulosa cells cultured under challenged conditions provide a model that can be used to examine substances for their ability to alter estradiol production and apoptosis in bovine granulosa cells. Mol. Reprod. Dev. 50:170–177, 1998. © 1998 Wiley‐Liss, Inc.

show abstract

Steroid production, cell proliferation, and apoptosis in cultured bovine antral and mural granulosa cells: Development of an in vitro model to study estradiol production

et al. 1998

View full text Add to dashboard Cite

This study was undertaken to characterize the relationship between changes in steroid production, cell cycle activity (ie, cell proliferation) and apoptosis in antral and mural bovine granulosa cells cultured in vitro. This was done to select conditions promoting optimal estradiol production by bovine granulosa cells cultured in completely defined conditions. In the first experiment, antral granulosa cells were cultured over the entire 4 days of the culture period in the presence of either 0, 2, or 10 ng/ml of FSH (chronic conditions) or were maintained under minimal FSH support (0.5 ng/ml FSH) for the first 3 days of culture and then were challenged over the fourth day of culture with either 0, 2, or 10 ng/ml FSH (challenged conditions). Compared with cells exposed to constant FSH levels (chronic conditions), the FSH-induced production of estradiol was higher (P < 0.006) and that of progesterone was lower (P < 0.02) over the last 24 h of culture, when antral granulosa cells were maintained under minimal FSH support during the first 3 days of culture (challenged conditions). In the second experiment, dynamics of estradiol and progesterone productions, conversion of [14C]androstenedione into subsequent steroid metabolites, DNA content, cell cycle activity, and apoptosis (as assessed by flow cytometry) of antral and mural granulosa cells over the first 3 days of culture under minimal FSH support and in response to a challenge with FSH during the last 24 h of culture were evaluated. Estradiol production as well as the conversion of androstenedione into testosterone and estradiol were greater (P < 0.01) in antral than in mural granulosa cells cultured under challenged conditions. A higher proportion of mural than antral granulosa cells were in the proliferative state at the end of culture (P < 0.03). This may be related to the decreased ability of mural cells to produce estradiol. FSH suppressed (P < 0.05) the spontaneous onset of apoptosis in both cell types. These results suggest that functional differences between these two cell compartments need to be considered in studying bovine granulosa cells in vitro. Because of their large (400 to 600%) FSH-induced estradiol production, antral granulosa cells cultured under challenged conditions provide a model that can be used to examine substances for their ability to alter estradiol production and apoptosis in bovine granulosa cells.

show abstract

Follicle-stimulating hormone-induced estradiol and progesterone production by bovine antral and mural granulosa cells cultured in vitro in a completely defined medium.

Cited by 30 publications

References 0 publications

Effect of FSH and cell localization on dimeric inhibin-A secretion from bovine granulosa cells in culture

Effect of FSH and cell localization on dimeric inhibin-A secretion from bovine granulosa cells in culture

Steroid production, cell proliferation, and apoptosis in cultured bovine antral and mural granulosa cells: Development of an in vitro model to study estradiol production

Steroid production, cell proliferation, and apoptosis in cultured bovine antral and mural granulosa cells: Development of an in vitro model to study estradiol production

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