title: The role of protein kinase-C in gonadotropin-induced ovulation in the in vitro perfused rabbit ovary.
abstract: Tumor-promoting phorbol esters are believed to affect ovarian granulosa cell progesterone and prostaglandin (PG) production and possibly ovulation by activating protein kinase-C (PKC). The effects of phorbol esters and PKC inhibitors on ovulation, progesterone, and PG production were examined in an in vitro perfused rabbit ovary. The effect of tranexamic acid, an inhibitor of the conversion of plasminogen activator to plasmin, on phorbol ester-induced ovulation was also examined. Phorbol 12,13-dibutyrate (PdBU), a PKC stimulator, induced ovulation in a dose-related manner in the absence of gonadotropins (56%, 200 nM PdBU; 0%, 0 nM PdBU; P < 0.05). Perfusate progesterone levels were increased only after 600 nM PdBU treatment, and perfusate PGF2 alpha, PGE2, and 6-keto-PGF1 alpha were increased in a dose-dependent fashion (P < 0.05). Staurosporine, a potent inhibitor of the catalytic domain of PKC, and calphostin-C, a specific inhibitor of the diacylglycerol-binding region, inhibited hCG-induced ovulation in a dose-related manner. Gonadotropin-induced ovulation decreased from 73% without staurosporine to 19% with 1.0 microM staurosporine (P < 0.01). Calphostin-C reduced ovulatory efficiency from 60% to 24% (P < 0.01). However, neither inhibitor decreased progesterone or PGF2 alpha production by ovaries exposed to hCG. hCG-induced oocyte maturation was also unaffected by exposure to either staurosporine or calphostin-C. Tranexamic acid reduced phorbol ester-induced ovulatory efficiency from 67% to 37% (P < 0.05). These findings demonstrate that the calcium-dependent PKC pathway is instrumental in gonadotropin-mediated follicular rupture in the rabbit. Although PGs may play an important role in ovulation, they do not appear to be directly responsible for PKC-mediated follicular rupture.
authors.affiliation: Department of Gynecology and Obstetrics, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
authors.forename: G
authors.initials: G
authors.lastname: Kaufman
authors.forename: A M
authors.initials: AM
authors.lastname: Dharmarajan
authors.forename: Y
authors.initials: Y
authors.lastname: Takehara
authors.forename: C S
authors.initials: CS
authors.lastname: Cropp
authors.forename: E E
authors.initials: EE
authors.lastname: Wallach
mesh_terms: D000818:Animals; D016922:Cellular Senescence / Q000187:drug effects; D006063:Chorionic Gonadotropin / Q000494:pharmacology*; D005260:Female; D066298:In Vitro Techniques; D009865:Oocytes / Q000502:physiology; D010053:Ovary / Q000187:drug effects*; D010060:Ovulation; D010477:Perfusion; D015240:Phorbol 12,13-Dibutyrate / Q000494:pharmacology; D011374:Progesterone / Q000097:blood; D011453:Prostaglandins / Q000097:blood; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000502:physiology*; D011817:Rabbits; D014148:Tranexamic Acid / Q000494:pharmacology
chemical_list: D006063:Chorionic Gonadotropin; D011453:Prostaglandins; D015240:Phorbol 12,13-Dibutyrate; D011374:Progesterone; D014148:Tranexamic Acid; D011493:Protein Kinase C