Periovulatory changes in steroid C17,20-lyase activity in ovaries of immature rats treated with pregnant mare serum gonadotropin.

It has been established that the effect of LH on ovarian steroid formation results initially in a rise of overall steroid production, which 2-4 h later is followed by a decrease in the formation of C19-steroids. To determine the reason for this decrease, the course of 17 alpha-hydroxyprogesterone (17 alpha-OHP) metabolism in immature rat ovaries (10,000 X g supernatant) stimulated to ovulate with pregnant mare serum gonadotropin (PMSG) was investigated. Employing optimal conditions the C17,20-lyase was estimated from the formation of androstenedione from 17 alpha-OHP as substrate. The Michaelis-Menten constant Km was 61 +/- 4 microM and V varied according to the time after PMSG administration. The initial activity at 0 h was 340 pmol min-1 mg-1 protein. It decreased 48 h after PMSG to 160 pmol, followed by a slight rise at 54 h and followed by a 60% fall at 57 h, at the height of the LH-surge. The activity declined to undetectable levels at 60 and 64 h (8 h before ovulation) and remained undetectable after ovulation at 72 and 96 h after PMSG. The enzymes 5 alpha-reductase and 20 alpha-hydroxysteroid dehydrogenase were estimated from the formed 3 alpha, 17 alpha-dihydroxy-5 alpha-pregnan-20-one and 17 alpha, 20 alpha-dihydroxy-4-pregn-en-3-one, respectively. In the unstimulated ovary the 5 alpha-reductase was 405 pmol; it decreased to 110 pmol 48 h after PMSG and continued to decrease. The 20 alpha-hydroxysteroid dehydrogenase activity which was undetectable in untreated rats, rose to 330 pmol 60 h after PMSG, 8 h before ovulation. The changes in enzyme activities when tracer amounts of [3H] 17 alpha-OHP were used agreed with the pattern of kinetic studies. From these incubations eight metabolites were isolated, one of them, 5 alpha-pregnane-3 alpha,17 alpha, 20 alpha-trioi, has not been previously identified. It is concluded that the decrease in C17,20-lyase activity limit the formation of C19-steroids in preovulatory follicles shortly before ovulation.