Hsp90의 저해제인 17-AAG의 처리에 따른 소 수정란의 배발달 및 세포사멸 양상

서지정보

ㆍ발행기관 : 한국동물번식학회 ㆍ수록지정보 : Reproductive & developmental biology / 35권 / 3호
ㆍ저자명 : 홍주희, 민성훈, 이에녹, 손형훈, 박흠대, 구덕본

목차

ABSTRACT 서론 재료 및 방법 배양액 체외성숙, 수정 및 배양 배반포에서 이중 형광 염색 TUNEL(Terminal Deoxynucleotidyl Transferase-MediateddUTP Nick-end Labeling) 분석 RNA 합성과 Real-time Quantitative PCR 통계학적 분석 결과 및 고찰 17-AAG 처리에 따른 소 수정란의 체외 발달 빛 배반포의 질적 수준 분석 소 배반포에서 세포사멸 양상과 관련 유전자의 발현에 있어서 17-AAG의 효과 결론 인용문헌

영어 초록

Heat shock protein 90 (Hsp90) is ATPase-directed molecular chaperon and affects survival of several cells. In our previous study, inhibitory effect of Hsp90 by inducing cell cycle arrest and apoptosis in the pig embryonic and primary cells was reported. However, its role during early bovine embryonic development is not sufficient. In this study, we traced the effects of Hsp90 inhibitor, 17-allylamino-17-demethoxygeldanamycin (17-AAG), on early bovine embryonic development. We also investigated several indicators of developmental potential, including structural integrity, gene expression (apoptosis-related genes), and apoptosis, which are affected by 17-AAG. Bovine embryos were cultured in the CR1-aa medium with or without 17-AAG for 7 days. In result, significant differences in developmental potential were detected between the embryos that were cultured with or without 17-AAG (33.1±9.6 vs 21.7± 8.3%). The structural integrity of the blastocysts was examined by differential staining. Blastocysts from the dbcAMP- treated group had higher numbers of ICM, TE, and total cells than those from the untreated group. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) showed that the number of containing fragmented DNA at the blastocyst stage increased in the 17-AAG treated group compared with control (11.2 vs 3.9, respectively). Blastocysts that developed in the 17-AAG treated group had low structural integrity and high apoptotic nuclei than those of the untreated control, resulting in decrease the embryonic qualities of preimplantation bovine blastocysts. The m-RNA expression of the pro-apoptotic gene (Bax) increased in 17-AAG treated group, whereas expression of the antiapoptotic gene (Bcl-XL) decreased. In conclusion, Hsp90 also appears to play a direct role in bovine early embryo developmental competence including structural integrity of blastocysts. Also, these results indicate that Hsp90 is closely associated with apoptosis-related genes expression in developing bovine embryos.

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