Characterization and regulation of a specific cell membrane receptor for somatomedin-C/insulin-like growth factor I in cultured rat granulosa cells.
The ovarian granulosa cell has recently been found to be a site of somatomedin-C/insulin-like growth factor I (Sm-C/IGF-I) production, reception, and action, thereby raising the prospect of a novel autocrine control mechanism concerned with granulosa cell ontogeny. It is the objective of the in vitro studies reported herein to explore the characteristics of the murine granulosa cell membrane Sm-C/IGF-I receptor and its regulation by gonadotropic, lactogenic, and beta 2-adrenergic signalling. Provision of FSH (150 ng/ml) to granulosa cells from immature rats cultured for 72 h under serum-free conditions resulted in a 3.1-fold increase over control values in specific cell-bound [125I]iodo-Sm-C/IGF-I. Binding to FSH-primed cells proved time, temperature, and pH dependent; optimal steady state conditions were achieved after an 8-h incubation at 15 C and a pH of 8.0. Although subject to regulation by the cellular density of plating, the binding of [125I]iodo-Sm-C/IGF-I to its receptor proved saturable (apparent Kd = 3.3 X 10(-9) M) as well as reversible; complete or partial tracer displacement was effected by competitive inhibition and dilution, respectively. Specificity studies revealed the competition for [125I]iodo-Sm-C/IGF-I binding to follow a relative rank order of potency of Sm-C/IGF-I much greater than multiplication-stimulating activity greater than insulin, but disclosed limited or no displacement by a series of chemically related and unrelated polypeptides. By Scatchard and Hill analysis, both control and FSH-treated cells displayed a single class of noninteracting binding sites; the FSH-enhanced binding represented largely increased binding capacity, rather than affinity. Significantly, up-regulation of granulosa cell Sm-C/IGF-I binding was not limited to FSH; qualitatively comparable increments in [125I]iodo-Sm-C/IGF-I binding were obtained after treatment with luteotropic, and beta 2-adrenergic (but not lactogenic) granulosa cell agonists. Taken together, these studies provide further evidence for the existence of high affinity, low capacity, specific cell membrane receptors for Sm-C/IGF-I in cultured rat granulosa cells. Our findings further indicate that the ability of FSH to enhance granulosa cell Sm-C/IGF-I binding largely reflects increased binding capacity rather than affinity and that this heterologous up-regulatory phenomenon may not be limited to FSH. As such, our observations of comparable up-regulation after luteotropic and beta 2-adrenergic (but not lactogenic) stimulation are in keeping with the view that cAMP may play an intermediary role in the regulation of granulosa cell type I IGF receptors.
D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D005260:Female; D006107:Granulosa Cells / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007334:Insulin-Like Growth Factor I / Q000378:metabolism*; D007700:Kinetics; D051381:Rats; D011919:Rats, Inbred Strains; D011972:Receptor, Insulin / Q000378:metabolism*; D017451:Receptors, Somatomedin; D013002:Somatomedins / Q000378:metabolism*
D017451:Receptors, Somatomedin; D013002:Somatomedins; D007334:Insulin-Like Growth Factor I; D011972:Receptor, Insulin