Porcine follicular fluid containing water-soluble LH/hCG receptor

Abstract

Porcine follicular fluid has been shown to have a specific water-soluble receptor for human chorionic gonadotropin (hCG). The binding of [125I] hCG to follicular fluid is inhibited by unlabelled hCG, LH but not FSH, ACTH and GH. The binding of hormone to the receptor in follicular fluid is a saturable phenomenon and Scatchard analysis suggested that the receptor has high affinity to hCG with no changes as the follicle enlarges. In contrast, follicular fluid from large follicles (6-12 mm) has higher binding capacity (2.04 +/- 0.12 fmol/mg protein) than follicular fluid isolated from medium (3-5 mm) and small (1-2 mm) follicles (0.60 +/- 0.05 and 0.44 +/- 0.04 fmol/mg protein, respectively). With the aid of affinity chromatography on hCG-CNBr-Sepharose 6-B a homogeneous fraction with Mr about 65,000 as estimated by SDS-PAGE was isolated. Treatment of follicular fluid with several protein-modifying reagents changed interactions of [125I] hCG with both soluble receptor and that bound to granulosa cell membrane in the similar manner. The [125I] hCG binding capacity of follicular fluid represents about 9.5% of the total binding capacity of granulosa cells. Finally, soluble LH/hCG receptor is probably secreted actively by follicular cells into follicular fluid. Dead granulosa cells do not release receptor into follicular fluid or incubation medium.