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Survivin protein expression in bovine follicular oocytes and their in vitro developmental competence
Authors:Jeon Kilsoo  Kim Eun Young  Tae Jin Cheol  Lee Chang Hyun  Lee Keum Sil  Kim Yeon Ok  Jeong Dong Kee  Cho Somi K  Kim Jae Hoon  Lee Hyo Yeon  Riu Key Zung  Cho Ssang Goo  Park Se Pill
Institution:aMirae Biotech./Cheju National University Stem Cell Research Center, Seoul 143-854, South Korea;bDepartment of Animal Biotechnology, Konkuk University, Seoul 143-702, South Korea;cDepartment of Animal Sciences, Chungbuk National University, Cheongju, Chungbuk 361-763, South Korea;dFaculty of Biotechnology, College of Applied Life Sciences, Cheju National University, 66 Jejudaehakno, Jeju 690-756, South Korea
Abstract:This study examined the relationship between survivin expression and the stage of development of in vitro cultured bovine oocytes and embryos; and whether survivin expression is affected by the quality of cumulus–oocyte complexes (COCS) or the quality of pre-implantation embryos. A polyclonal antibody was prepared using recombinant bovine survivin protein. Expression of survivin mRNA and protein was analyzed by real-time quantitative RT-PCR and immunocytochemistry. In the first experiment, survivin mRNA expression was examined at developmental stages from germinal vesicle (GV) oocyte to blastocyst, it was significantly decreased after fertilization of matured oocytes (P < 0.05), then increased slightly to the 8-cell stage followed by rapid increases at the morula and blastocyst stages (P < 0.05). In the second experiment, the effect of oocyte quality on survivin protein, pro-apoptotic (bax, caspase-3) and anti-apoptotic (survivin, bax inhibitor) mRNA expression was examined. Survivin protein was more strongly expressed in good quality COCS than in poor quality COCS. The expression of the anti-apoptotic genes, survivin and bax inhibitor, was significantly higher (P < 0.05) and that of the pro-apoptotic genes, bax and caspase-3, was significantly lower (P < 0.05) in good compared to poor quality COCS. The developmental competence of good quality COCS (30.4% blastocysts) was significantly better than that of poor quality COCS. In the last experiment also, we confirmed that significantly higher expression of survivin and bax inhibitor genes and significantly lower expression of bax and caspase-3 genes was resulted in good quality blastocysts than in poor quality blastocysts (P < 0.05). It was concluded that the expression of survivin was related to the quality of COCS, their developmental competence and the quality of in vitro produced blastocysts. Consequently, survivin may be a good candidate marker for embryo quality.
Keywords:Bovine  In vitro  Embryo  Survivin  Apoptotic genes
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