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대한생식의학회지 제24권 제1호 2010년

체내에서 성숙이 재개된 생쥐난자의 투명대 경화

서울여자대학교 자연과학대학 생물학과;서울여자대학교 자연과학대학 생물학과;피엘산부인과 체외수정연구실;피엘산부인과 체외수정연구실;충북대학교 자연과학대학 생물학과;한양대학교 자연과학대학 생물학과;

김지수;김해권;박종민;이승재;이준영;김문규;,

Zona Hardening of Mouse Oocytes Undergone Meiotic Resumption In Vivo

Kim, Ji-Soo;Kim, Hae-Kwon;Park, Jong-Min;Lee, Seung-Jae;Lee, Joon-Young;Kim, Moon-Kyoo;

Department of Biology, Seoul Women's University;Department of Biology, Seoul Women's University;IVF Research Laboratory, Park & Lee Infertility Clinic;IVF Research Laboratory, Park & Lee Infertility Clinic;Department of Biology, Choongbuk University;Department of Biology, Hanyang University;

It is well known that the zona pellucidae of mouse oocytes become "hardened" when they are allowed to mature in vitro in the absence of serum components. To see if oocytes already undergone meiotic resumption in vivo exhibit similar zona hardening, hardening of ZP of cumulus-enclosed oocytes(CEOs) was examined after culture in vitro since their release from follicles various hours after hCG injection. When CEOs matured in vivo for 3h or longer were subjected to culture in vitro for 14h with BSA alone, zona hardening was significantly reduced compared to those cultured in vitro from the begining of maturation. However, when CEOs matured in vivo for 5h were freed from cumulus cells and then cultured in vitro with BSA alone, little reduction of zona hardening was observed. Preincubation of CEOs for 5h with fetuin, one of the well known inhibitor of in vitro zone hardening, did not prevent zona hardening during its subsequent culture of CEOs for 14h without fetuin. However, when CEOs precultured with both fetuin and PMSG for 5h and then further cultured with BSA alone for 14h, zona hardening was dramatically reduced. Under these conditions, the expansion of cumulus cell was observed. In addition, CEOs cultured with both BSA and dbcAMP to prevent their meiotic resumption showed a significant increase of zona hardening. Whether the observed zona hardening was correlated with the conversion of ZP2 to $ZP2_{f}$ was examined. Zona pellucida, isolated from CEOs matured for 5h in vivo and then further cultured with BSA alone was subjected to SDS-PAGE. Most of ZP2 molecules from these CEOs did not undergo conversion from ZP2 to $ZP2_{f}$. From these results, it is concluded that CEOs undergone meiotic resumption in vivo do not exhibit zona hardening when they were subsequently cultured in vitro without serum components. It appears that cumulus cells play an important role in this phenomenon.

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