Abstract
To evaluate the relation between the steroidogenic activity and cell proliferation of individual follicles in mature hamster ovaries during the estrous cycle, the localization of enzymes involved in estrogen biosynthesis and bromodeoxyuridine (BrdU) incorporation were examined immunohistochemically. Moreover, granulosa cells from the early atretic follicle were examined by scanning and transmission electron microscopy. Immunoreactivity for aromatase was localized in the granulosa cells of healthy developing follicles and Graafian follicles, as well as in newly formed granulosa lutein cells. In the healthy follicles of an ovulation cycle, intensity of aromatase immunoreactivity was suddenly decreased on day 3. The theca interna cells of healthy developing follicles were immunopositive for 17α-hydroxylase/C17–C20 lyase (17α-lyase) from day 2 to the morning of day 4, but on the evening of day 4 most theca interna cells were immunonegative except for only a few cells of the large Graafian follicles. BrdU incorporation was observed in the granulosa cells of healthy developing follicles, in the endothelial cells of capillaries around the developing follicles, and of newly formed corpora lutea. Very early morphological signs of atresia was the pyknotic change of a few granulosa cells lining the antral cavity. In that follicle, the number of BrdU-incorporating granulosa cells was suddenly decreased whilst immunoreactivity of aromatase and 17α-lyase were gradually decreased. These data suggest that the mechanism of the loss of aromatase activity from the granulosa cells of atretic follicles appears to differ from that in cycling follicles. Even in the early stage of atresia, the granulosa cells showed remarkable morphological change characteristic of apoptosis, as visualized by scanning and transmission electron microscopy. Cessation of granulosa cell proliferation may occur earlier than apoptotic change and the degeneration of the granulosa cells becomes rapid once atresia starts.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bill CH, Gilbert SG (1981) Acute gonadotropin deprivation. 1. A model for the study of follicular atresia. Biol Reprod 24: 913–921
Byskov AG (1979) Atresia. In: Midgley AR, Sadler WA (eds) Ovarian follicular development and function. Raven Press, New York, pp 41–57
Carson RS, Findlay JK, Clarke IJ, Burger HG (1981) Estradiol, testosterone, and androstenedione in ovine follicular fluid during growth and atresia of ovarian follicles. Biol Reprod 24: 105–113
de Fazio A, Leary JA, Hedley DW, Tattersall MHN (1987) Immunohistochemical detection of proliferating cells in vivo. J Histochem Cytochem 35: 571–577
Hay MF, Cran DG, Moor RM (1976) Structural changes occurring during atresia in sheep ovarian follicles. Cell Tissue Res 169: 515–529
Hirshfield AN (1989) Rescue of atretic follicles in vitro and in vivo. Biol Reprod 40: 181–190
Hughes FM Jr, Gorospe WC (1991) Biochemical identification of apoptosis (programmed cell death) in granulosa cells: evidence for a potential mechanism underlying follicular atresia. Endocrinology 129: 2415–2422
Inoue T, Osatake H (1988) A new drying method of biological specimens for scanning electron microscopy. Arch Histol Jpn 51: 53–59
Ishimura K, Yoshinaga-Hirabayashi T, Tsuri H, Fujita H, Osawa Y (1989) Further immunocytochemical study on the localization of aromatase in the ovary of rats and mice. Histochemistry 90: 413–416
Ishimura K, Yoshinaga-Hirabayashi T, Tsuri H, Kominami S, Takemori S, Fujita H (1990) Immunocytochemical and biochemical studies on the localization and changes of 17α-hydroxylase/C17–C20 lyase activity in immature rat ovary treated with PMSG and hCG. Histochemistry 94: 225–229
Knigge KM, Leathem JH (1956) Growth and atresia of follicles in the ovary of the hamster. Anat Rec 124: 680–707
Kominami S, Shinzawa K, Takemori S (1982) Purification and some properties of cytochrome P-450 specific for steroid 17α-hydroxylation and c17–c20 bond cleavage from guinea pig adrenal microsomes. Biochem Biophys Res Commun 109: 916–921
Le Goascogne C, Sananes N, Gouezou M, Baulieu EE, Robel P (1989) Cell-specific variations and hormonal regulation of immunoreactive cytochrome P-450SCC in rat ovary. J Reprod Fertil 85: 61–72
Maxson WS, Haney AF, Schomberg DW (1985) Steroidogenesis in porcine atretic follicles: loss of aromatase activity in isolated granulosa and theca. Biol Reprod 33: 495–501
Osawa Y, Higashiyama T, Fronckowiak N, Yoshida N, Yarborough C (1987a) Aromatase. J Steroid Biochem 27: 781–789
Osawa Y, Yoshida N, Fronckowiak M, Kitawaki J (1987b) Immunoaffinity purification of aromatase cytochrome P-450 from human placental microsomes, metabolic switching from aromatization to 1β and 2β monohydroxylation, and recognition of aromatase isozyme. Steroids 50: 11–28
Sasano H, Okamoto M, Mason JI, Simpson ER, Mendelson CR, Sasano N, Silverberg SG (1989) Immunolocalization of aromatase, 17α-hydroxylase and side-chain-cleavage cytochromes P-450 in the human ovary. J Reprod Fertil 85: 163–169
Shinzawa K, Ishibashi S, Murakoshi M, Watanabe K, Kominami S, Kawahara A, Takemori S (1988) Relationship between zonal distribution of microsomal cytochrome P-450s (P-45017α lyase, and P-450c21) and steroidogenic activities in guinea pig adrenal cortex. J Endocrinol 119: 191–200
Terranova PF (1981) Steroidogenesis in experimentally induced atretic follicles of hamster: a shift from estradiol to progesterone synthesis. Endocrinology 108: 1885–1890
Tilly JL, Kowalski KI, Schomberg DW, Hsueh AJW (1992) Apoptosis in atretic ovarian follicles is associated with selective decreases in messenger ribonucleic acid transcripts for gonadotropin receptors and cytochrome P450 aromatase. Endocrinology 131: 1670–1676
Tsuri H, Yoshinaga-Hirabayashi T, Ishimura K, Osawa Y, Kominami S, Takemori S, Fujita H (1992) Immunocytochemical studies on the localization of aromatase and 17α-hydroxylase/C17–20-lyase (17α-lyase) in estrous cycling and pregnant hamster ovaries. Arch Histol Cytol 55: 13–20
Uilenbroek JThJ, Woutersen PJA, van der Schoot P (1980) Atresia of preovulatory follicles: gonadotropin binding and steroidogenic activity. Biol Reprod 23: 219–229
Wyllie AH (1980) Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature 284: 555–556
Wyllie AH, Kerr JR, Currie AR (1980) Cell death: the significance of apoptosis. Int Rev Cytol 68: 251–306
Yoshinaga-Hirabayashi T, Ishimura K, Fujita H, Inano H, Ishii-Ohba H, Tamaoki B (1987) Immunocytochemical localization of 17β-hydroxysteroid dehydrogenase (17β-HSD), and relation to the ultrastructure of steroidogenic cells in immature and mature rat ovaries. Arch Histol Jpn 50: 545–556
Yoshinaga-Hirabayashi T, Ishimura K, Fujita H, Kitawaki J, Osawa Y (1990) Immunocytochemical localization of aromatase in immature rat ovaries treated with PMSG and hCG, and in pregnant rat ovaries. Histochemistry 93: 223–228
Zhong C, Ishimura K, Yoshinaga-Hirabayashi T, Fujita H, Kitawaki J, Osawa Y (1989) Immunocytochemical study on the localization of aromatase in the ovary of golden hamster, guinea pig and cow. Acta Histochem Cytochem 22: 501–507
Zlotkin T, Farkash Y, Orly J (1986) Cell-specific expression of immunoreactive cholesterol side-chain cleavage cytochrome P-450 during follicular development in the rat ovary. Endocrinology 119: 2809–2820
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Yoshinaga-Hirabayashi, T., Osawa, Y. Steroidogenic activity of atretic follicles in the cycling hamster ovary and relation to ultrastructural observations. Histochemistry 102, 59–67 (1994). https://doi.org/10.1007/BF00271050
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00271050