Role of apoptosis on human follicular growth and analyzes of regulattry mechanism of the apoptosis in granulosa cells

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1) We analyzed apotosis by TUNEL method in the human granulosa cells obtained when in vitro fertilization was performed. The granulosa cells were cultured, and time-couse of apoptosis occurrence were analyzed with or without hCG in the culture media, however, some samples showed apoptosis and the others showed no apoptosis throughout the course. No effects of hCG were observed. These results indicated that the apoptosis in the granulosa or luteal cells might not involved the regulation of cell turn-over in these cells.2) To clarify whether apoptosis is involved endometriosis, we obtained eutopic endometrial tissues along with endometriotic tissues from the adenomyosis in 12 cases and from the ovary in 12 other cases. Apoptosis-induced DNA fragmentation was detected by the TUNEL method, and immunostaining with a monoclonal antibody against the Fas, Le^y or bcl-2 was also performed using the same tissue section. Analysis showed that apoptosis was involved in all the ovarian endometriotic tissues but in only 2 of the 12 adenomyotic tissues and in 5 of the 24 eutopic endometrial tissues. None of these cases correlated to phases of the menstrual cycle. The expression of bcl-2 in the eutopic endometrial and adenomyotic tissues was limited to be proliferative phase, and was observed in only one of the 12 cases of ovarian endometriosis. Fas and Le^y were expressed randomly across a wide range in both the eutopic and ectopic endometrial tissues. These results suggest that features of ovarian endometriosis is defferent from adenomyosis and eutopic endometrium in terms of apoptosis involvement.