Survivin protein expression in bovine follicular oocytes and their in vitro developmental competence.
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.
Jeon
Kilsoo
K
Mirae Biotech./Cheju National University Stem Cell Research Center, Seoul 143-854, South Korea.
Kim
Eun Young
EY
Tae
Jin Cheol
JC
Lee
Chang Hyun
CH
Lee
Keum Sil
KS
Kim
Yeon Ok
YO
Jeong
Dong Kee
DK
Cho
Somi K
SK
Kim
Jae Hoon
JH
Lee
Hyo Yeon
HY
Riu
Key Zung
KZ
Cho
Ssang Goo
SG
Park
Se Pill
SP
D000818:Animals; D001755:Blastocyst / Q000502:physiology*; D015153:Blotting, Western / Q000662:veterinary; D053148:Caspase 3 / Q000737:chemistry / Q000235:genetics; D002417:Cattle / Q000196:embryology / Q000235:genetics / Q000502:physiology*; D003001:Cloning, Molecular; D047108:Embryonic Development / Q000502:physiology*; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D018507:Gene Expression Regulation, Developmental; D007150:Immunohistochemistry / Q000662:veterinary; D008869:Microtubule-Associated Proteins / Q000096:biosynthesis* / Q000235:genetics; D009865:Oocytes / Q000502:physiology*; D011247:Pregnancy; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D051028:bcl-2-Associated X Protein / Q000737:chemistry / Q000235:genetics
D008869:Microtubule-Associated Proteins; D012333:RNA, Messenger; D011994:Recombinant Proteins; D051028:bcl-2-Associated X Protein; D053148:Caspase 3