The specific binding of [125I]iodo human chorionic gonadotropin to porcine granulosa cells isolated at two stages of follicle maturation was quantitated immediately after harvest and following 6-7 days of culture. Both porcine LH (pLH, LER 786-3) and hCG inhibited 125I]iodo hCG binding, while pFSH (LER-1132) did not compete for hCG binding sites. The incubation of labeled hCG with granulosa cells for 24-48 hours at 37 C did not alter its subsequent ability to bind to fresh cells. The binding of [125I]iodo hCG was a rapid process which was not readily reversible. Scatchard analysis of the equilibrium binding data resulted in linear plots showing the hCG binding capacity of highly differentiated (HD) cells, harvested from large preovulatory follicles, to be significantly greater than that of moderately differentiated (MD) cells isolated from smaller luteal phase follicles, both before (794vs 93 pmoles/g; P < 0.001) and after (157 vs 5 pmoles/g; P < 0.001) culture. A decline in binding capacity occurred in both cell groups during culture and was associated with a significant decrease in progestin production. In contrast, the hCG binding affinity of the granulosa cell was equivalent at both stages of follicle development and remained unchanged after 6 days of culture (mean apparent Kd = 2.9 × 10-10M; n = 27). These data indicate that porcine granulosa cells contain a homogeneous class of LH receptors whose number, but not affinity, increases during follicle maturation in vivo. The loss of receptors in vitro suggests the absence of some factor(s), as yet unidentified, critical to the maintenance and development of the receptor population.
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