Activin-A inhibits progesterone production by macaque luteal cells in culture

John D. Brannian, Teresa K. Woodruff, Jennie P. Mather, Richard Stouffer

    Research output: Contribution to journalArticle

    27 Citations (Scopus)

    Abstract

    Since inhibin is produced during the luteal phase of the menstrual cycle in women and nonhuman primates, the primate corpus luteum (CL) may be a local site of inhibin/activin action. This study was designed to determine whether inhibin or activin altered steroidogenesis by macaque luteal cells in vitro. Luteal cells were obtained by enzymatic dispersion of CL from rhesus monkeys at midluteal phase of the menstrual cycle. Cells (2 × 104/0.2 mL) were cultured in wells coated with extracellular matrix from bovine corneal endothelial cells in Dulbecco's modified Eagle's medium/F-12 medium (1:1 vol/vol) + insulin (2 ng/mL), transferrin (5 ng/mL), H2SeO3 (0.25 nmol), and aprotinin (10 μg/mL). Various concentrations (0-400 ng/mL) of recombinant human-inhibin-A, recombinant human-activin-A or human CG (hCG) (100 ng/mL; CR123) alone or in combination with inhibin or activin were added to the culture media (n = 5 Exp). Media were changed daily for 4 days and progesterone (P) concentrations were determined by RIA. Inhibin exposure did not alter P levels compared to that of control (untreated) cultures. In contrast, activin (10-400 μg/ mL) suppressed P production (P <0.05) below controls and inhibin-treated cultures by days 3 and 4. Exposure to hCG increased P levels throughout culture (9 x control levels by day 4; P <0.05). hCG-stimulated P production was unaltered by inhibin, whereas activin (50-400 ng/mL) reduced (maximal inhibition of 40%; P <0.05) hCG-stimulated P production by day 4 of culture. Cell number on day 4 was not altered by any dose of inhibin or activin, but the number of cells staining for 3β-hydroxysteroid dehydrogenase was reduced (P <0.05) by 32.9 ± 2.6% in activin-treated cultures. Since P levels declined during culture in all treatment groups, in a second series of experiments (n = 4), luteal cells were cultured for 4 days with or without hCG (100 ng/mL) and low density lipoprotein (LDL; 100 μg/mL) ± 0-400 ng activin/mL. P production in the presence of hCG + LDL was greatly enhanced compared to other treatment groups and was sustained during days 2-4 of culture. Activin at doses of 50-400 ng/mL suppressed (maximal inhibition of ≈35%; P <0.05) hCG + LDL-stimulated P production on days 3 and 4. These results suggest that the primate CL is a target for activin action to suppress luteal cell activities, including gonadotropin-regulated, lipoprotein-mediated steroidogenesis.

    Original languageEnglish (US)
    Pages (from-to)756-761
    Number of pages6
    JournalJournal of Clinical Endocrinology and Metabolism
    Volume75
    Issue number3
    StatePublished - Sep 1992

    Fingerprint

    Luteal Cells
    Activins
    Macaca
    Inhibins
    Progesterone
    Cell Culture Techniques
    Corpus Luteum
    Primates
    3-Hydroxysteroid Dehydrogenases
    Cell Count
    activin A
    Eagles
    Aprotinin
    Luteal Phase
    Level control
    Endothelial cells
    Transferrin
    Menstrual Cycle
    Macaca mulatta
    Gonadotropins

    ASJC Scopus subject areas

    • Biochemistry
    • Endocrinology, Diabetes and Metabolism

    Cite this

    Activin-A inhibits progesterone production by macaque luteal cells in culture. / Brannian, John D.; Woodruff, Teresa K.; Mather, Jennie P.; Stouffer, Richard.

    In: Journal of Clinical Endocrinology and Metabolism, Vol. 75, No. 3, 09.1992, p. 756-761.

    Research output: Contribution to journalArticle

    Brannian, John D. ; Woodruff, Teresa K. ; Mather, Jennie P. ; Stouffer, Richard. / Activin-A inhibits progesterone production by macaque luteal cells in culture. In: Journal of Clinical Endocrinology and Metabolism. 1992 ; Vol. 75, No. 3. pp. 756-761.
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    abstract = "Since inhibin is produced during the luteal phase of the menstrual cycle in women and nonhuman primates, the primate corpus luteum (CL) may be a local site of inhibin/activin action. This study was designed to determine whether inhibin or activin altered steroidogenesis by macaque luteal cells in vitro. Luteal cells were obtained by enzymatic dispersion of CL from rhesus monkeys at midluteal phase of the menstrual cycle. Cells (2 × 104/0.2 mL) were cultured in wells coated with extracellular matrix from bovine corneal endothelial cells in Dulbecco's modified Eagle's medium/F-12 medium (1:1 vol/vol) + insulin (2 ng/mL), transferrin (5 ng/mL), H2SeO3 (0.25 nmol), and aprotinin (10 μg/mL). Various concentrations (0-400 ng/mL) of recombinant human-inhibin-A, recombinant human-activin-A or human CG (hCG) (100 ng/mL; CR123) alone or in combination with inhibin or activin were added to the culture media (n = 5 Exp). Media were changed daily for 4 days and progesterone (P) concentrations were determined by RIA. Inhibin exposure did not alter P levels compared to that of control (untreated) cultures. In contrast, activin (10-400 μg/ mL) suppressed P production (P <0.05) below controls and inhibin-treated cultures by days 3 and 4. Exposure to hCG increased P levels throughout culture (9 x control levels by day 4; P <0.05). hCG-stimulated P production was unaltered by inhibin, whereas activin (50-400 ng/mL) reduced (maximal inhibition of 40{\%}; P <0.05) hCG-stimulated P production by day 4 of culture. Cell number on day 4 was not altered by any dose of inhibin or activin, but the number of cells staining for 3β-hydroxysteroid dehydrogenase was reduced (P <0.05) by 32.9 ± 2.6{\%} in activin-treated cultures. Since P levels declined during culture in all treatment groups, in a second series of experiments (n = 4), luteal cells were cultured for 4 days with or without hCG (100 ng/mL) and low density lipoprotein (LDL; 100 μg/mL) ± 0-400 ng activin/mL. P production in the presence of hCG + LDL was greatly enhanced compared to other treatment groups and was sustained during days 2-4 of culture. Activin at doses of 50-400 ng/mL suppressed (maximal inhibition of ≈35{\%}; P <0.05) hCG + LDL-stimulated P production on days 3 and 4. These results suggest that the primate CL is a target for activin action to suppress luteal cell activities, including gonadotropin-regulated, lipoprotein-mediated steroidogenesis.",
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    N2 - Since inhibin is produced during the luteal phase of the menstrual cycle in women and nonhuman primates, the primate corpus luteum (CL) may be a local site of inhibin/activin action. This study was designed to determine whether inhibin or activin altered steroidogenesis by macaque luteal cells in vitro. Luteal cells were obtained by enzymatic dispersion of CL from rhesus monkeys at midluteal phase of the menstrual cycle. Cells (2 × 104/0.2 mL) were cultured in wells coated with extracellular matrix from bovine corneal endothelial cells in Dulbecco's modified Eagle's medium/F-12 medium (1:1 vol/vol) + insulin (2 ng/mL), transferrin (5 ng/mL), H2SeO3 (0.25 nmol), and aprotinin (10 μg/mL). Various concentrations (0-400 ng/mL) of recombinant human-inhibin-A, recombinant human-activin-A or human CG (hCG) (100 ng/mL; CR123) alone or in combination with inhibin or activin were added to the culture media (n = 5 Exp). Media were changed daily for 4 days and progesterone (P) concentrations were determined by RIA. Inhibin exposure did not alter P levels compared to that of control (untreated) cultures. In contrast, activin (10-400 μg/ mL) suppressed P production (P <0.05) below controls and inhibin-treated cultures by days 3 and 4. Exposure to hCG increased P levels throughout culture (9 x control levels by day 4; P <0.05). hCG-stimulated P production was unaltered by inhibin, whereas activin (50-400 ng/mL) reduced (maximal inhibition of 40%; P <0.05) hCG-stimulated P production by day 4 of culture. Cell number on day 4 was not altered by any dose of inhibin or activin, but the number of cells staining for 3β-hydroxysteroid dehydrogenase was reduced (P <0.05) by 32.9 ± 2.6% in activin-treated cultures. Since P levels declined during culture in all treatment groups, in a second series of experiments (n = 4), luteal cells were cultured for 4 days with or without hCG (100 ng/mL) and low density lipoprotein (LDL; 100 μg/mL) ± 0-400 ng activin/mL. P production in the presence of hCG + LDL was greatly enhanced compared to other treatment groups and was sustained during days 2-4 of culture. Activin at doses of 50-400 ng/mL suppressed (maximal inhibition of ≈35%; P <0.05) hCG + LDL-stimulated P production on days 3 and 4. These results suggest that the primate CL is a target for activin action to suppress luteal cell activities, including gonadotropin-regulated, lipoprotein-mediated steroidogenesis.

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