Prostaglandin production and ovulation during exposure of amphibian ovarian follicles to gonadotropin or phorbol ester in vitro.
Experiments were carried out at different times of hibernation to ascertain whether prostaglandin is produced by Rana ovarian follicles during gonadotropin- or 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced in vitro ovulation. Ovarian fragments were cultured in amphibian Ringer in the presence or absence of frog pituitary homogenate (FPH, 0.05 gland/ml) or TPA (1 or 10 microM). After various periods of culture, incidence of ovulation was determined and prostaglandin F2 alpha (PGF2 alpha) accumulated in culture medium was measured by radioimmunoassay. FPH and TPA increased PGF2 alpha levels in medium in a dose-dependent manner. The time course of PGF2 alpha secretion and ovulation by FPH or TPA treatment varied during the hibernation period. In early-hibernation, FPH stimulated neither PGF2 alpha secretion nor ovulation while TPA stimulated PGF2 alpha secretion, although it failed to induce ovulation. In mid-hibernation, FPH and TPA effectively stimulated PGF2 alpha secretion and ovulation, but both events took place several hours later than those observed in late-hibernation. Some fragments obtained in mid-hibernation and most obtained in late-hibernation spontaneously produced PGF2 alpha in vitro without FPH or TPA treatment and in some instances spontaneously ovulated. Furthermore, FPH or TPA increased PGF2 alpha levels further or accelerated the time course of secretion by such fragments. In late-hibernation, PGF2 alpha secretion induced with FPH or TPA increased simultaneously with or later than onset of ovulation. Exogenous cAMP (2.5 mM) or 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine (H-7, 100 microM), a PKC inactivator, markedly suppressed FPH- or TPA-stimulated PGF2 alpha secretion and ovulation in mid-hibernation. Indomethacin (IM, 5 micrograms/ml) strongly suppressed TPA- or FPH-stimulated PGF2 alpha production by fragments but its effect on ovulation varied among animals and at different times. IM suppressed ovulation of some ovarian fragments obtained in mid-hibernation, but failed to suppress hormone-induced ovulation in late-hibernation. Cycloheximide (5 micrograms/ml) and actinomycin D (1 microgram/ml) effectively suppressed FPH-stimulated PGF2 alpha production and ovulation, whereas actinomycin D reduced but failed to significantly suppress TPA-induced PGF2 alpha production in mid-hibernation. In general, effects of ovulation inhibitors exhibited strong seasonal variations and were less efficient as the breeding season approached. Taken together, the data suggest that elevated levels of PGF2 alpha are associated with spontaneous and hormone-induced ovulation, and PKC mediates gonadotropin induction of PGF2 alpha but not steroid synthesis in Rana ovaries.
Chang
K J
KJ
Department of Biology, Chonnam National University, Kwangju, Republic of Korea.
Kim
J W
JW
Lee
J
J
Im
W B
WB
Kwon
H B
HB
Schuetz
A W
AW
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / Q000494:pharmacology; D000818:Animals; D002273:Carcinogens / Q000494:pharmacology*; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000494:pharmacology; D003513:Cycloheximide / Q000494:pharmacology; D016861:Cyclooxygenase Inhibitors / Q000494:pharmacology; D003609:Dactinomycin / Q000494:pharmacology; D015237:Dinoprost / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D006062:Gonadotropins / Q000494:pharmacology*; D007213:Indomethacin / Q000494:pharmacology; D010060:Ovulation / Q000187:drug effects / Q000378:metabolism*; D010703:Phorbol Esters / Q000494:pharmacology*; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D011863:Radioimmunoassay; D011898:Ranidae
D002273:Carcinogens; D016861:Cyclooxygenase Inhibitors; D004791:Enzyme Inhibitors; D006062:Gonadotropins; D010703:Phorbol Esters; D011500:Protein Synthesis Inhibitors; D003609:Dactinomycin; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D003513:Cycloheximide; D015237:Dinoprost; D000242:Cyclic AMP; D007213:Indomethacin