Molecular and Cellular Endoerfnoiogy 5 (1976) 0 North-Holland Publishing Company
A SYNERGiSTIC
315-320
EFFECT OF ANDROGEN ON THE STIMULATION
PROGESTERONE CELLS
OF
SECRETION BY FSH IN CULTURED RAT GRANULOSA
A. NIMR~D and H.R. LINDNER Department
of Hormone Research, The Weizmann Institute of‘ Science, Rehovot, Israel
Received 31 March, 1976
Gcanulosa cells from preovulatory follicles (PO) or from the enlarged preantral follicles of hypophysectomized immature diethylstilbestrol-treated (Hx-DES) rats were cultured with various combinations of FSH, androst4_ene-3,17dione (Ad), estradioL17P and dibutyryl cyclic AMP (dbcAMP). Progestin levels (progesterone and 20Gdihydroprogesterone) in the medium after 2 days of culture were assayed by radio~munossay. The control levels of the two progestins were lower for Hx-DES than for PO cells. Rat FSH ~~AMD-I-3; 0.1 &ml) caused a 2fold rise in progestin accumulation in both PO and Hx-DES cultures. dbcAMP (1 mM) increased progestin accumulation in PO cultures 445-fold, and to an even greater extent (lo-20 fold) in Hx-DES cuttures. Androstenedione (1.0 Mg/ml) augmented progestin accumulation (IS-3-fold), and synergized the steroidogenic action of FSH: in cells from Fix-DES rats, combined treatment with FSH and Ad caused a S-lo-fold increase over the values obtained with FSH alone. Testosterone and Sadihydrotestosterone, but not estradiot-17p or estrone, mimicked these effects of Ad. Ad did not synergize the action of dbcAMP on progestin levels in HxDES cultures. It is proposed that androgen may play a role in the development of the FSHresponsive mechanism in preantral granulosa cells. Keywords:
20a-dihydroprogesterone; androstenedione; testosterone; Sa-dihydrotestosterone; cyclic AMP; follicular maturation; androgen; FSH; progesterone secretion, granulosa cell.
Follicle-stimulating hormone (FSH) has been shown to play an important role in follicular development (Hisaw, 1947; Eshkol and Lunenfeld, 1972). The granulosa cells of the ovarian follicle are endowed with FSH receptors (Midgley 1973; Presl et al, 1974; Nimrod et al., 1976), and addition of FSH to cultured granulosa cells results in increased secretion of progestins (Channing, 1974; Erickson, Nimrod and Ryan, unpublished). The effect of steroid hormones on follicular development is less clear. Granulosa cells have been shown to possess cytoplasmic receptors for estrogens (Richards, 1975) and androgens (Schreiber and Ross, 1975). While estrogen may play a part in the induction of the LH receptor by FSH (Richards and Midgley, 1976), no definite role has yet been assigned to androgens in follicular de-
316
A. Nimrod, H.R. Lindner
velopment, though both mitogenic and anti-mitogenic effects on granulosa cells have been reported (Gaarenstroom and DeJongh, 1946; Louvet et al., 1975). We therefore examined the effect of androgens on the stimulation of progestin secretion by FSH in cultured rat granulosa cells.
EXPERIMENTAL
PROCEDURE
Granulosa cells were collected either from mature rats on the morning (9 to 10 a.m.) of proestrus (PO-cells) or from immature hypophysectom~~ed rats treated with diethylstilbestrol (0.5 mg/rat/day) for 5 days (Hx-DES cells; Nimrod et al., 1976) and cultured as described by Erickson et al. (1974), except that 5% serum from hypophysectomized rats was included in the medium instead of rabbit serum. Rat FSH (NIAMD-I-3, LH contamination
A SYNERGiSTIC
315-320
EFFECT OF ANDROGEN ON THE STIMULATION
PROGESTERONE CELLS
OF
SECRETION BY FSH IN CULTURED RAT GRANULOSA
A. NIMR~D and H.R. LINDNER Department
of Hormone Research, The Weizmann Institute of‘ Science, Rehovot, Israel
Received 31 March, 1976
Gcanulosa cells from preovulatory follicles (PO) or from the enlarged preantral follicles of hypophysectomized immature diethylstilbestrol-treated (Hx-DES) rats were cultured with various combinations of FSH, androst4_ene-3,17dione (Ad), estradioL17P and dibutyryl cyclic AMP (dbcAMP). Progestin levels (progesterone and 20Gdihydroprogesterone) in the medium after 2 days of culture were assayed by radio~munossay. The control levels of the two progestins were lower for Hx-DES than for PO cells. Rat FSH ~~AMD-I-3; 0.1 &ml) caused a 2fold rise in progestin accumulation in both PO and Hx-DES cultures. dbcAMP (1 mM) increased progestin accumulation in PO cultures 445-fold, and to an even greater extent (lo-20 fold) in Hx-DES cuttures. Androstenedione (1.0 Mg/ml) augmented progestin accumulation (IS-3-fold), and synergized the steroidogenic action of FSH: in cells from Fix-DES rats, combined treatment with FSH and Ad caused a S-lo-fold increase over the values obtained with FSH alone. Testosterone and Sadihydrotestosterone, but not estradiot-17p or estrone, mimicked these effects of Ad. Ad did not synergize the action of dbcAMP on progestin levels in HxDES cultures. It is proposed that androgen may play a role in the development of the FSHresponsive mechanism in preantral granulosa cells. Keywords:
20a-dihydroprogesterone; androstenedione; testosterone; Sa-dihydrotestosterone; cyclic AMP; follicular maturation; androgen; FSH; progesterone secretion, granulosa cell.
Follicle-stimulating hormone (FSH) has been shown to play an important role in follicular development (Hisaw, 1947; Eshkol and Lunenfeld, 1972). The granulosa cells of the ovarian follicle are endowed with FSH receptors (Midgley 1973; Presl et al, 1974; Nimrod et al., 1976), and addition of FSH to cultured granulosa cells results in increased secretion of progestins (Channing, 1974; Erickson, Nimrod and Ryan, unpublished). The effect of steroid hormones on follicular development is less clear. Granulosa cells have been shown to possess cytoplasmic receptors for estrogens (Richards, 1975) and androgens (Schreiber and Ross, 1975). While estrogen may play a part in the induction of the LH receptor by FSH (Richards and Midgley, 1976), no definite role has yet been assigned to androgens in follicular de-
316
A. Nimrod, H.R. Lindner
velopment, though both mitogenic and anti-mitogenic effects on granulosa cells have been reported (Gaarenstroom and DeJongh, 1946; Louvet et al., 1975). We therefore examined the effect of androgens on the stimulation of progestin secretion by FSH in cultured rat granulosa cells.
EXPERIMENTAL
PROCEDURE
Granulosa cells were collected either from mature rats on the morning (9 to 10 a.m.) of proestrus (PO-cells) or from immature hypophysectom~~ed rats treated with diethylstilbestrol (0.5 mg/rat/day) for 5 days (Hx-DES cells; Nimrod et al., 1976) and cultured as described by Erickson et al. (1974), except that 5% serum from hypophysectomized rats was included in the medium instead of rabbit serum. Rat FSH (NIAMD-I-3, LH contamination
