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Synthetic luteinizing hormone-releasing hormone (LH-RH) lost both its immunoreactivity and hormonal activity on incubation with hypothalamic or cerebrocortical slices or homogenates. This inactivation was shown to be due to degradation of the decapeptide by soluble enzyme(s) present in the 100,000 x g supernatant fraction of the homogenates. The supernatant derived from one rat hypothalamus was capable of destroying 1 /.L g of exogenous LH-RH within 5 min. The hexapeptide pGlu-His-Trp-Ser-Tyr-Gly was identified as the major radioactive breakdown product of [ pGl~-3-~H] LH-RH, and tentative evidence for the formation of the tetrapeptide Leu-Arg-Pro-Gly-NH2 was obtained by sequential electrophoresis and paper chromatography. These findings suggest that the Gly-Leu bond may be the preferred site of cleavage.

Reference preparations of ovine follicle-stimulating hormone (NIH-FSH-S8 and S9; 10-50 mug/ml) induced ovum maturation and stimulated cyclic AMP formation, as well as progesterone and 17beta-estradiol secretion, by rat Graafian follicles in vitro. These actions of NIH-FSH were retained after immunoabsorption of any contaminating luteinizing hormone (LH) present in the preparations, by treatment with an antiserum to the beta-subunit of purified ovine LH (anti-betaLH). In contrast, the corresponding biological actions of NIH-LH-S18 (0.5-10 mug/ml) were abolished by treatment with this anti-betaLH serum. A highly purified FSH preparation (64-96 CD, 0.25 mug/ml) also triggered oocytic meiosis and increased follicular progesterone secretion in vitro. Intraperitoneal (ip) administration of anti-betaLH-treated NIH-FSH-S9 (50 mug/rat at 1430 h) consistently induced ovulation in proestrous rats in which the endogenous gonadotropin surge had been blocked by ip injection of either Nembutal (1345 h) or antiserum to the LH-releasing hormone (1200 h). Injection (ip) of anti-betaLH serum on its own into proestrous rats at 1200 h prevented ovum maturation and follicular rupture. We conclude that currently available reference preparations of ovine FSH possess the capacity to stimulate follicular adenylate cyclase, steroidogenesis, and ovum maturation in vitro, as well as ovulation in vivo, in the rat, and that this capacity cannot be attributed to contamination with material immunochemically identical with LH. However, it is inferred that the physiological triggering of ovulation and related events in this species depends principally on LH.

The intravenous administration of prostaglandin E₂ (PGE₂; 100 µg/rat) to immature male and female rats increased the serum level of luteinizing hormone (LH). This effect was prevented by the prior administration of an antiserum to the hypothalamic gonadotropin-releasing hormone. LH release from rat pituitaries in vitro was not stimulated by PGE₂ (10 µg/ml). It is inferred that the stimulatory effect of PGE₂, on LH release is not a direct one on the pituitary gland, but is exerted at the level of the hypothalamus.