Kondo H, Maruo T, Peng X, Mochizuki M
Department of Obstetrics and Gynecology, Kobe University School of Medicine, Japan.
J Clin Endocrinol Metab. 1996 Jul;81(7):2702-10. doi: 10.1210/jcem.81.7.8675599.
Immunohistochemical localization of the Fas antigen in the infant and adult human ovary during follicular growth, regression, and atresia was examined by the avidin/biotin immunoperoxidase method with a monoclonal antibody to the Fas antigen. Western blotting was used to confirm the presence of the Fas antigen protein. In primordial and primary follicles within the normal adult ovary, only the oocyte showed moderate immunostaining for the Fas antigen. In secondary and antral follicles, only the oocyte showed weak staining for the Fas antigen, while in preovulatory follicles, neither the oocyte nor the granulosa and theca cells were immunostained for the Fas antigen. In corpora lutea, the Fas antigen staining became apparent in the granulosa lutein cells during the early luteal phase and intensified during the mid luteal phase, while the theca lutein cells became positive for the Fas antigen staining during the mid luteal phase. During the late luteal phase, the staining intensity of the Fas antigen in the regressing corpora lutea further increased. As the regressing corpora lutea were converted into corpora albicans, the staining intensity decreased, and the corpora albicans and stromal cells were negative for the Fas antigen. In atretic primordial and primary follicles, only the degenerating oocyte showed the Fas antigen staining. By contrast, in atretic antral follicles, the Fas antigen staining was profound in the degenerating granulosa cells at the early stage of atresia, and at the mid stage of atresia it was intensified in the cell surface of the scattered granulosa cells and became apparent in the theca cells. At the late stage of atresia the Fas antigen remained only in the hypertrophied theca cells. In the infant ovary, only the oocyte in primordial and primary follicles exhibited intense staining for the Fas antigen. In the postmenopausal ovary, the Fas antigen staining was entirely negative. Western blot analysis revealed the presence of the Fas antigen protein with a molecular mass of 45 kDa in luteal tissues. On the basis of the recent evidence, that the Fas antigen mediates an apoptotic signal in a variety of cells, the abundant expression of the Fas antigen in the regressing corpora lutea and atretic follicles suggests that the Fas antigen participates in luteal regression and follicular atresia through the apoptotic process. Furthermore, notable expression of the Fas antigen in the oocyte of primordial and primary follicles within the infant and adult human ovary followed by the decrease in the Fas antigen expression in the oocyte with the advance of follicular maturation suggests that the Fas antigen expression in the oocyte may play a role in follicular selection.
采用抗Fas抗原单克隆抗体,运用抗生物素蛋白/生物素免疫过氧化物酶法,检测了Fas抗原在婴幼儿及成年女性卵巢卵泡生长、退化和闭锁过程中的免疫组织化学定位。利用蛋白质印迹法证实Fas抗原蛋白的存在。在正常成年卵巢的原始卵泡和初级卵泡中,仅卵母细胞对Fas抗原呈中度免疫染色。在次级卵泡和有腔卵泡中,仅卵母细胞对Fas抗原呈弱阳性染色,而在排卵前卵泡中,卵母细胞、颗粒细胞和卵泡膜细胞均未被Fas抗原免疫染色。在黄体中,Fas抗原染色在黄体早期的颗粒黄体细胞中开始显现,并在黄体中期增强,而卵泡膜黄体细胞在黄体中期开始呈现Fas抗原染色阳性。在黄体后期,退化黄体中Fas抗原的染色强度进一步增加。随着退化黄体转变为白体,染色强度降低,白体和基质细胞对Fas抗原呈阴性。在闭锁的原始卵泡和初级卵泡中,仅退化的卵母细胞显示Fas抗原染色。相比之下,在闭锁的有腔卵泡中,闭锁早期退化的颗粒细胞中Fas抗原染色明显,闭锁中期,散在颗粒细胞的细胞表面染色增强,并在卵泡膜细胞中显现。在闭锁后期,Fas抗原仅存在于肥大的卵泡膜细胞中。在婴幼儿卵巢中,仅原始卵泡和初级卵泡中的卵母细胞对Fas抗原呈强阳性染色。在绝经后卵巢中,Fas抗原染色完全阴性。蛋白质印迹分析显示,黄体组织中存在分子量为45 kDa的Fas抗原蛋白。基于最近的证据,即Fas抗原在多种细胞中介导凋亡信号,Fas抗原在退化黄体和闭锁卵泡中的大量表达表明,Fas抗原通过凋亡过程参与黄体退化和卵泡闭锁。此外,Fas抗原在婴幼儿及成年女性卵巢原始卵泡和初级卵泡的卵母细胞中显著表达,随后随着卵泡成熟进程卵母细胞中Fas抗原表达降低,提示卵母细胞中Fas抗原的表达可能在卵泡选择中发挥作用。