Identification of Gonadotropin Surge-Inhibiting Factor (GnSIF) in Follicular Fluid and its Differentiation from Inhibin1

Abstract

Exposure of rat pituitary cell cultures to charcoal-extracted porcine follicular fluid (pFF) inhibits gonadotropin-releasing hormone (GnRH) stimulation of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) release in a dose-dependent manner. The inhibition of GnRH-stimulated gonadotropin secretion (gonadotropin surge-inhibiting factor [GnSIF] activity) by pituitary cells in vitro requires up to 24-48 h preexposure to pFF. One microliter of pFF inhibits approximately 50% of the GnRH-stimulated LH release and is defined as 1 unit of GnSIF activity. Basal LH secretion is unaltered under these conditions. GnSIF activity is distinct from that of inhibin, which selectively suppresses basal release of FSH but not LH. A partially purified preparation of inhibin contains less than 1% as much GnSIF activity as inhibin activity. GnSIF activity is resistant to moderate heat treatment (60 degrees C for 60 min) and is fully recovered after acetone precipitation. Chromatography of pFF on heparin/Sepharose affinity matrix effectively separates inhibin from GnSIF. Whereas inhibin has a high affinity for heparin, GnSIF activity does not associate with this affinity matrix and is recovered in the column void volume. In summary, we have developed an in vitro bioassay for the detection of GnSIF activity in pFF. Moreover GnSIF activity seemingly derives from a molecular entity distinct from inhibin, having different physicochemical characteristics and differential effects on pituitary gonadotropin secretion.