Abstract

Using high-performance liquid chromatography and a combination of electrochemical and radiometric flow detection for 2-[14C]hydroxyestradiol, changes in estrogen 2-hydroxylase activity in the microsomal fraction of rat ovarian homogenates were followed. Injection of human chorionic gonadotropin (hCG) at 12-hr intervals to hypophysectomized immature rats stimulated hypertrophy of the theca-interstitial tissue and produced a profound increase in enzyme activity. With the last injection of hCG at 96 hr the peak serum concentration of hCG was reached 12 hr later and then decreased exponentially with a half-time of 13 hr. However, enzyme activity remained elevated for at least 60 hr before beginning to fall. Pregnant mare's serum gonadotropin (PMSG) also produced an increase in activity, which was apparently limited to the thecal-interstitial tissue because freshly removed granulosa cells from the mature follicles had undetectable activity levels. Administration of anti-PMSG antiserum after enzyme activity had been increased resulted in a prompt fall in activity, as did injection of hCG to mimic an ovulatory surge of LH. The results indicate that the thecal-interstitial tissue of the rat ovary has estrogen 2-hydroxylase activity that is dependent upon gonadotropic stimulation for expression.

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