EFFECTS OF INSULIN-LIKE GROWTH FACTOR-I ON FOLLICLE GROWTH, OOCYTE MATURATION, AND OVARIAN STEROIDOGENESIS AND PLASMINOGEN ACTIVATOR ACTIVITY IN THE RABBIT

摘要

We examined the effects of insulin-like growth factor (IGF)-I on follicular growth, oocyte maturation, and ovarian steroidogenesis and plasminogen activator (PA) activity in vitro, using a perfused rabbit ovary preparation in order to determine whether the follicle-stimulating effects of growth hormone (CH) are mediated by IGF-I. The addition of IGF-I to the perfusate stimulated follicular growth and the resumption of meiosis in follicular oocytes in a dose-dependent manner. There was no significant difference in the production of progesterone by perfused rabbit ovaries between IGF-I-treated and control ovaries, whereas ICF-I increased the production of estradiol (E(2)) by perfused rabbit ovaries in a dose-dependent manner. The concomitant addition of a monoclonal antibody recognizing the type I ICF receptor, alpha IR-3, to the perfusate significantly blocked IGF-I-stimulated follicular growth, oocyte maturation, and E(2) production. Intrafollicular PA activity increased significantly 4 h after exposure to 10 or 100 ng/ml of IGF-I and reached maximal levels at 6 h. The percentage increase in follicle diameter at 6 h after exposure to ICF-I was significantly correlated with the intrafollicular PA activity. Treatment with Ctl resulted in a 2.7-fold increase in intrafollicular levels of ICF-I mRNA. The binding of [I-125]-IGF-I to rabbit ovarian membrane preparations was inhibited by unlabeled IGF-I and ICF-II in a concentration-dependent manner. The relative affinity of the IGF-I receptor for IGF-I, ICF-II, and insulin was typical of type 1 binding (IGF-I > IGF-II > insulin). Affinity cross-linking of ovarian membranes with [I-125]-IGF-I revealed a radiolabeled band corresponding to a molecular weight of 135 000, the alpha subunit of the type I IGF receptor. This band was totally displaced by IGF-I and alpha IR-3. It was concluded that IGF-I stimulated follicular development, E(2) production, and oocyte maturation by interacting with its specific receptor located in rabbit ovarian membranes.