Metabolism of steroid hormones In vitro by the theca and granulosa layer of the ovarian follicle in the hen.
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The homogenates of the theca layer and granulosa layers of the ovarian follicle of 300-day-old White Leghorn hens, obtained 6 hours and 1-0.3 hour before the predicted ovulation, were incubated at 41°C with 14C-labelled steroids in the presence of NAD+or NADPH. After one hour incubation, radioactive steroid metabolites were extracted and analyzed by thin-layer chromatography.When radioactive pregnenolone was used as the substrate, progesterone was obtained as the major metabolite by the granulosa cells, but not by the theca cells. The amount of the metabolite was higher in the granulosa cells of the largest follicle (F1) than the third largest follicle (F3). When radioactive progesterone was used as the substrate, 17 α-hydroxyprogesterone was obtained as the major metabolite by the theca cells, but not by the granulosa cells. The amount of the metabolite was much higher in the theca cells of F3 than F1. When radioactive 17 α-hydroxyprogesterone was used as the substrate, androstenedione and testosterone were produced as the major metbolites by the theca cells. The total amount of the metabolites tended to be higher in the cells of F3 than F1. When radioactive androstenedione was used as the substrate, testosterone was obtained as the major metabolite in considerable amounts by both theca cells and granulosa cells either of F1 or F3.These results indicate that the combined activities of Δ5 steroid-3β-hydroxysteroid dehydrogenase and Δ5-Δ4 isomerase, estimated from the amount of progesterone, were extremely high in the granulosa cells of F1, and high in those of F3, but very low in the theca cells either of F1 or F3; and that the activity of 17 α-hydroxylase, estimated with the amount of 17 α-hydroxyprogesterone, was extremely high in theca cells of F3, and high in those of F1, but very low in the granulosa cells either of F1 or F3. The activity of C17-20 lyase, estimated from the amount of androstenedione, was high in the theca cells of F3, whereas the activity of 17 α-hydroxysteroid dehydrogenase, estimated the amount of testosterone, was high both in the theca cells and granulosa cells either of F1 or F3. The results well account for very high concentration of progesterone in the granulosa cells of F1, and higher amounts of testosterone and estradiol in the theca cells of F3 and more smaller follicles of the hen.
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