Summary
A systematic study of Δ5-3β-hydroxysteroid dehydrogenase in the granulosa cells of immature and mature mice was made. The histochemical results were compared with the ultrastructural findings on the same cells in an attempt to determine whether the granulosa cells are capable of a steroidogenic role.
In newborn and immature mice the granulosa cells of a great amount of follicles demonstrated a moderate or strong histochemical activity. In mature mice the granulosa cells demonstrated a weak or moderate activity normally only in preovulatory follicles and in some other atretic follicles. The granulosa cells of the normal developing follicles did not show such activity. In addition the histological control of numerous parallel sections demonstrated particularly in immature ovaries the presence of a great amount of atretic follicles.
In the cytoplasm of the granulosa cells of the follicles in immature ovaries only clusters of lipid droplets with ribosomes were noted; while in the preovulatory follicles of mature animals there started to appear mitochondria with tubular cristae, smooth membranes of the endoplasmic reticulum and irregular lipid droplets. In the obviously atretic follicles several granulosa cells as well as theca interna cells showed numerous lipid droplets and ribosomes together with different degenerating organelles. The granulosa cells of the normal developing follicles showed a well developed Golgi complex, granular endoplasmic reticulum and free ribosomes.
The histochemical and ultrastructural findings suggest a steroidogenic role of the granulosa cells only in the larger preovulatory follicles (probably related to an early luteinization of this layer) but this role was not demonstrated in the same cells in normal developing follicles.
In addition, since an histochemical positivity was demonstrated also in the granulosa cells of some obviously atretic follicles, it is possible that many of the follicles having granulosa cells filled with lipid droplets and attached ribosomes and histochemically positive might be, in the immature ovaries, in a very precocious stage of atresia.
It is to precise for these cells whether a cytoplasm with these two strictly correlated components (lipid droplets and attached ribosomes) and showing an histochemical positivity could carry-on all the biochemical steps involved in steroid biosynthesis or only has only a temporary capability to produce some precursors of steroids.
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References
Bjersing, L.: On the ultrastructure of follicles and isolated follicular granulosa cells of porcine ovary. Z. Zellforsch. 82, 173–186 (1967A).
Bjersing, L.: Histochemical demonstration of Δ5-3β- and 17β-hydroxysteroid dehydrogenase activities in porcine ovary. Histochemie 10, 295–304 (1967B).
Bjersing, L., Carstensen, H.: The role of the granulosa cell in biosynthesis of ovarian steroid hormones. Biochim. biophys. Acta (Amst.) 86, 639–640 (1964).
Björkman, N.: A study of the ultrastructure of the granulosa cells of the rat ovary. Acta anat. (Basel) 51, 125–147 (1962).
Blanchette, E.J.: Ovarian steroid cells. I. Differentiation of the lutein cells from the granulosa follicle cell during the preovulatory stage and under the influence of exogenous gonadotrophins. J. Cell Biol. 31, 501–516 (1966).
Botte, V., Del Bianco, C.: Research in the histochemical distribution of lipids and steroid 3β-ol-dehydrogenase in the ovaries of immature rats treated with gonadotrophins. Arch. Ostet. Ginec. 67, 653–665 (1962).
Brambell, F.W.R.: Ovarian changes. In: Marshall's physiology of reproduction, vol. I, part 1. A. S. (Parkes, ed.), p. 397–542. London: Longmans Green and Co. 1956.
Christensen, A.K., Gillim, S.W.: The correlation of fine structure and function in steroid-secreting cells, with emphasis on those of the gonads. In: The gonads, (McKerns, ed.), chap. 16, p. 415–488. New York: A.C.C., 1969.
Dahl, E.: The fine structure of the granulosa cells in the domestic fowl and the rat. Z. Zellforsch. 119, 58–67 (1971).
Flerkó, B., Hajós, F., Sétáló, G.Y.: Electron microscopic observations on rat ovaries in different stages of development and steroidogenesis. Acta morph. Acad. Sci. hung. 15, 163–183 (1967).
Franceschini, M.P., Santoro, A., Motta, P.: L'ultrastruttura delle cellule della granulosa nelle varie fasi di maturazione del follicolo ooforo. Z. Anat. Entwickl.-Gesch. 124, 522–532 (1965).
Franchi, L.L., Mandl, A.M.: The ultrastructure of oogonia and oocytes in the foetal and neonatal rat. Proc. roy. Soc. 157, 99–114 (1962).
Gondos, B.: The ultrastructure of granulosa cells in the newborn rabbit ovary. Anat. Rec. 165, 67–78 (1969).
Hadek, R.: Electron microscope study on primary liquor folliculi secretion in the mouse ovary. J. Ultrastruct. Res. 9, 445–458 (1963).
Hadjioloff, A.I.: Contribution à l'histologie théorique. I. Du tissu sanguin et du tissu sexuel. Ann. Univ. Fac. Med. Sofia 11, 457–472 (1932).
Hadjioloff, A.I.: Manuel d'histologie et d'embryologie, Medizina et Fysk. Sofia (1967).
Hadjioloff, A.I., Bojadjieva-Michailova, A.: Histochemische und elektronenmikroskopische Untersuchungen der Lipide im Eierstock von Mäuseembryonen und neugeborenen weißen Mäusen. In: A.I. Hadjioloff (ed.), Histochimie et cytochimie des lipides. V. Symposium Internat. des Histologistes à Sofia, Edit. Acad. Bulg. Sci. 179–186 (1966).
Hart, D.McK., Baillie, A.H., Calman, K.C., Ferguson, M.M.: Hydroxysteroid dehydrogenase development in the mouse adrenal and gonads. The ovary. In: Developments in steroid histochemistry, chap. 5, p. 72–102. London and New York: Acad. Press. 1966.
Levy, H., Deane, H.W., Rubin, B.L.: Visualization of steroid 3β-ol-dehydrogenase activity in tissues of intact and hypophysectomized rats. Endocrinology 65, 932–943 (1959).
Luft, J.H.: Improvements in epoxy resin embedding methods. J. biophys. biochem. Cytol. 9, 409–414 (1961).
Millonig, G.: Advantages of a phosphate buffer for OsO4 solutions in fixation. J. appl. Physiol. 32, 1637 (1961).
Motta, P.: Sulla formazione del liquor folliculi nell'ovaio della coniglia. Biol. lat. (Milano) 18, 252–271 (1965).
Motta, P.: Histochemical evidence of early stages of atretic follicles in different mammals. In: T. Takeuchi, K. Ogawa, S. Fujita (eds.). 4th International Congress of Histochemistry and Cytochemistry, August, Kyoto, pp. 599–600, 1972.
Müller, H.G., Linnartz-Niklas, A.: Autoradiographische Untersuchung über die Größe der Eiweiß-Synthese der weiblichen Genitalorgane im Metoestrus bei Ratte und Maus. Arch. Gynäk. 194, 48–62 (1960).
Odor, D.L.: The ultrastructure of unilaminar follicles of the hamster ovary, Amer. J. Anat. 116, 493–521 (1965).
Odor, D.L., Blandau, R.J.: Ultrastructural studies on fetal and early postnatal mouse ovaries. I. Histogenesis and organogenesis. Amer. J. Anat. 124, 163–186 (1969A).
Odor, D.L., Blandau, R.J.: Ultrastructural studies on fetal and early postnatal mouse ovaries. II. Cytodifferentiation. Amer. J. Anat. 125, 177–216 (1969B).
Presl, J., Jirasek, J., Horsky, J., Henzel, M.: Observations on steroid 3β-ol-dehydrogenase activity in the ovary during early postnatal development in the rat. J. Endocr. 31, 293–294 (1965).
Reynolds, E.S.: The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J. Cell Biol. 17, 208–212 (1963).
Rubin, B.L., Deane, H.W., Hamilton, J.A., Driks, E.: Changes in Δ5-3β-hydroxysteroid dehydrogenase activity in the ovaries of maturing rats. Endocrinology 72, 924–930 (1963).
Stegner, H.E., Wartenberg, H.: Elektronenmikroskopische und histotopochemische Untersuchungen über Struktur und Bildung der Zona pellucida menschlicher Eizellen. Z. Zellforsch 53, 702–712 (1961).
Watson, M.L.: Staining of tissue sections for electron microscopy with heavy metals. J. biophys. biochem. Cytol. 4, 475–478 (1958).
Wattenberg, L.W.: Microscopic histochemical demonstration of steroid 3β-ol-dehydrogenase in tissue sections. J. Histochem. Cytochem. 6, 225–232 (1958).
Watzka, M.: Weibliche Genitalorgane. Das Ovarium: Handbuch der mikroskopischen Anatomie des Menschen, Bd. VII, Tl. 3, S. 1–178. Berlin-Göttingen-Heidelberg: Springer 1957.
Weakley, B.S.: Electron microscopy of the oocyte and granulosa cells in the developing ovarian follicles of the golden hamster (Mesocricetus auratus). J. Anat. (Lond.) 100, 503–534 (1966).
Weakley, B.S.: Light and electron microscopy of developing germ cells and follicle cells in the ovary of the golden hamster: twenty-four hours before birth to eight days post partum. J. Anat. (Lond.) 101, 435–459 (1967).
Weakley, B.S.: Differentiation of the surface epithelium of the hamster ovary. An electron microscopic study. J. Anat. (Lond.) 105, 129–147 (1969).
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The present results were partially presented to the “56ème Congrès de l'Association des Anatomistes” (Nantes, 4–8/4/1971) and to the “66° Verhandlungen der Anatomischen Gesellschaft” (Zagreb, 2/4/1971).
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Hadjioloff, A.I., Bourneva, V. & Motta, P. Histochemical demonstration of Δ5-3 ß-OHD activity in the granulosa cells of ovarian follicles of immature and mature mice correlated with some ultrastructural observations. Z.Zellforsch 136, 215–228 (1973). https://doi.org/10.1007/BF00307442
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DOI: https://doi.org/10.1007/BF00307442