Summary
During the last four days of follicular development prior to ovulation, the activities of Δ 5-3β-hydroxysteroid dehydrogenase (3βOHD) and glucose-6-phosphate dehydrogenase (G-6-PD) were quantified in cryostat sections of the rat ovary. The product of the enzyme reactions were measured using a scanning and integrating microdensitometer. The enzyme activity was measured in the peripheral region, the antral region and the cumulus of the membrana granulosa (MG) of these follicles on the morning of each of the four days of the estrous cycle. G-6-PD activity was measured in the presence and absence of an intermediate hydrogen acceptor, phenazine methosulphate, to provide a measure of the quantity of Type I and Type II Hydrogen (H) generated: Type I H is considered to be related to hydroxylating reactions such as those of steroids and Type II H to other general biosynthetic activities of cells.
In all three regions of the MG of follicles of the ovulable type, 3βOHD activity was lowest in estrus and diestrus-1, increased on diestrus-2 and peaked in proestrus. In estrus and diestrus-1, the level of 3βOHD activity in the three regions was comparable. However, by diestrus-2, and even more conspicuously in proestrus, enzyme activity was significantly greater in the peripheral region than in the antral region or in the cumulus. During the same period, the level of enzyme activity remained comparable in the last two regions. Throughout the estrous cycle, both Type I and Type II H generation from G-6-PD was greatest in the peripheral region, less in the antral region and least in the cumulus. In the peripheral region, Type I H generation increased progressively after diestrus-1, to reach a maximum in proestrus. In the antral region, Type I H generation increased between diestrus-1 and diestrus-2 and then remained unchanged through proestrus. In the cumulus, Type I H generation remained at levels seen in estrus throughout the remainder of the cycle. Generation of Type II H, in the peripheral region was constant throughout the estrous cycle. In contrast, in the antral region and cumulus, Type II H generation was greater in diestrus-1 and diestrus-2 than on either proestrus or estrus.
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Armstrong, D.T.: Gonadotropins, ovarian metabolism and steroid biosynthesis. Rec. Progr. Horm. Res. 24, 255–319 (1968)
Bratt, H., Pupkin, M., Weisz, J., Lloyd, C.W., Balogh K., Jr.: Dehydrogenases in II. A histochemical study of steroid and carbohydrate metabolizing enzyme in micropolycystic ovaries induced by continuous illumination. Endocrinology 83, 329–335 (1968)
Channing, C.P.: The use of granulosa cell cultures and short-term incubations in the assay for gonadotropins. In: Gonadotropins and gonadal function. N.R. Mougdal (ed.), pp. 185–198. New York: Academic Press 1974
Chayen, J., Altman, F.P., Butcher R.G.: The effect of certain drugs on the production and possible utilization of reducing equivalents outside the mitochondria. In: Fundamentals of cell pharmacology. S. Dikstein, (ed.), pp. 196–230. Springfield, IL: Charles C. Thomas 1973a
Chayen, J., Bitensky, L., Butcher R.: Practical histochemistry. London: J. Wiley and Sons, 1973b
Cortes, V., McCracken, J.A., Lloyd, C.W., Weisz, J.: Progestin production by the ovary of the testosterone-sterilized rat treated with an ovulatory dose of LH, and the normal, proestrous rat. Endocrinology 89, 878–885 (1971)
Dixon, W.J. and F.J. Massey, Jr.: Introduction to statistical analysis. New York: McGraw Hill 1957
Falck B.: Site of production of oestrogen in rat ovary as studied in microtransplants. Acta Physiol. Scand. 47, (Suppl. 163), 5–101 (1959)
Makris, A., Ryan K.J.: Progesterone, androstenedione, testosterone, estrone and estradiol synthesis in hamster ovarian follicle cells. Endocrinology 96, 694–701 (1975)
McKerns, K.W.: Studies on the regulation of ovarian function by gonadotrophins. In: The Gonads. K.W. McKerns (ed.), pp. 137–174. New York: Appleton-Century-Crofts 1969
Prabhu, V.K.K., Weisz J: Effect of blocking ovulation in the rat by pentobarbital on ovarian 3β-hydroxysteroid dehydrogenases: A histochemical study. Endocrinology 87, 481–485 (1970)
Pupkin, M., Bratt, H., Weisz, J., Lloyd, C.W., Balogh K., Jr.: Dehydrogenases in the rat ovary. I. Histochemical study of Δ 5-3β-hydroxysteroid dehydrogenases and enzymes of carbohydrate oxidation during the estrous cycle. Endocrinology 79, 316–327 (1966)
Samuels, L.T., Helmreich, M.L., Lasater, M.B., Reich H.: An enzyme in endocrine tissues which oxidizes Δ 5-3β-hydroxysteroid to α,β-unsaturated ketones. Science 113, 490–491 (1951)
Short, R.V.: Steroids in the follicular fluid and the corpus luteum of the mare. A “two-cell type” theory of ovarian steroid synthesis. J. Endocrinol. 24, 59–63 (1962)
Stoklosowa, S., Nalbandov, A.V.: Luteinization and steroidogenic activity of rat ovarian follicles cultured in vitro. Endocrinology 91, 25–32 (1972)
Thompson, E.A., Siiteri, P.V.: Partial resolution of the placental microsomal aromatase complex. J. Steroid Biochem. 7, 635–639 (1976)
Yoshinaga, K., Hawkins, R., Stocker J.: Estrogen secreted by the rat ovary in vivo during the estrous cycle and pregnancy. Endocrinology 85, 103–112 (1969)
Fortune, J.E., Armstrong D.T. Hormonal control of 17β-estradiol biosynthesis in proestrous rat follicle: estradiol production by isolated theca versus granulosa. Endocrinology 102, 227–235 (1978)
Guraya, S.S.: Comparative studies on the histochemical features of ovarian compartments in the rat and golden hamster, with special reference to steroid hormone synthesis. Acta Anat. 82, 284–300 (1972)
Ham, A.W.: Histology. Philadelphia: J.B. Lippincott 1969
Hoyer, P.E., Andersen H.: Histochemistry of 3β-hydroxysteroid dehydrogenase in rat ovary. I. A. methodological study. Histochemistry 51, 167–193 (1977)
Juchau, M.R., Zachariah P.K.: Displacement of carbon monoxide from placental cytochrome P-450 by steroid: antagonistic effects of androstenedione and 19-norandrostenedione. Biochem. Biophys. Res. Commun. 65, 1026–1032 (1975)
Kidwell, W.R., Balogh, K., Jr., Weist W.G.: Effects of luteinizing hormone on glucose-6-phosphatase dehydrogenase activities in superovulated rat ovaries. Endocrinology 79, 352–361 (1966)
Kovarik, F.A., Kasprow, B.A., Emmert, L.A., Velardo J.T.: Localization of ovarian dehydroepian-drosterone and pregnenolone-3β-hydroxysteroid dehydrogenases in the rat during the estrous cycle. In: Biology of Reproduction, Basic and Clinical Studies. J.T. Velardo and B.A. Kasprow (eds.). 1971
Lindner, A., Amsterdam, Y., Salamon, A., Tsafriri, A., Nimrod, A., Lamprecht, S.A., Zor, U., Koch, Y.: Intraovarian factors in ovulation: determinant of follicular response to gonadotrophins. J. Reprod. Fertil. 51, 215–235 (1977)
Wiest, W.B., Kidwell W.R.: The regulation of progesterone secretion by ovarian dehydrogenases. In: The Gonads. K.W. McKerns (ed.), pp. 295–325. New York: Appleton-Century-Crofts 1969
Zoller, L.C., Weisz J.: The structure and acid phosphatase activity in granulosa cells of preovulatory follicles in rat ovary. Anat. Rec. 190, 592 (1978a)
Zoller, L.C., Weisz J.: Identification of cytochrome P-450 in the membrana granulosa of the preovulatory follicle, using quantitative cytochemistry. Endocrinology 103, 310–313 (1978b)
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This work was supported by research grants from the National Institute of Child Health and Human Development (# HD-12684) and (# HD-09542) and from the Rockefeller Foundation
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Zoller, L.C., Weisz, J. A quantitative cytochemical study of glucose-6-phosphate dehydrogenase and Δ 5-3β-hydroxysteroid dehydrogenase activity in the Membrana granulosa of the ovulable type of follicle of the rat. Histochemistry 62, 125–135 (1979). https://doi.org/10.1007/BF00493314
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DOI: https://doi.org/10.1007/BF00493314