The Effect of Ovary Storage and In Vitro Maturation on mRNA Levels in Bovine Oocytes; A Possible Impact of Maternal ATP1A1 on Blastocyst Development in Slaughterhouse-derived Oocytes
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概要
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Since BSE testing of slaughtered cattle is obligatory in Japan, storage of ovaries at 15-20 C overnight in phosphate buffered saline has become a routine protocol in in vitro production (IVP) of cattle embryos. Ovary storage is known to reduce developmental competence of oocytes; however, its effects on oocyte gene expression have not been clarified yet. This study compared oocytes collected from stored slaughterhouse-derived ovaries with those collected by Ovum Pick-Up (OPU) in terms of the expression of 20 selected genes to determine if ovary storage affects cellular processes at the molecular level. Expression of mRNA in oocytes was assayed before and after in vitro maturation (IVM) by real-time quantitative PCR. Maternal mRNA levels of genes were investigated in 2-cell stage embryos obtained from slaughterhouse oocytes to assess their roles for blastocyst formation. In immature OPU oocytes, genes related to metabolism (GAPDH), transporters (GLUT8, ATP1A1) and stress resistance protein (HSP70) showed significantly higher expression compared with oocytes derived from stored ovaries. During IVM, the expression of GDF9, GLUT8, CTNNB1 and PMSB1 was significantly decreased irrespective of oocyte source. Two-cell stage embryos cleaving at 22-25 h after in vitro fertilization (IVF) showed a significantly higher blastocyst formation rate and ATP1A1 gene expression level compared with those cleaving at 27-30 h after IVF. Our results reveal that storage of ovaries alters mRNA levels in oocytes. Correlation of Na/K ATPase ATP1A1expression in IVP embryos at the 2-cell and 8-cell stages with their developmental ability to the blastocyst stage may suggest the importance of maternal mRNA of this gene during blastulation in embryos derived from slaughterhouse oocytes.
著者
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Somfai Tamas
National Livestock Breeding Center
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Inaba Yasushi
National Livestock Breeding Center
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IMAI Kei
National Livestock Breeding Center
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WATANABE Shinya
National Institute of Livestock and Grassland Science
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Imai Kei
National Inst. Agrobiological Sci. Ibaraki Jpn
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Geshi Masaya
National Institute Of Livestock And Grassland Science
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AKAGI Satoshi
National Institute of Livestock and Grassland Science
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Nagai Takashi
National Inst. Of Livestock And Grassland Sci.
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KANEDA Masahiro
National Institute of Livestock and Grassland Science, National Agriculture and Food Research Organization, Ibaraki 305-0901, Japan
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HARAGUCHI Seiki
National Institute of Livestock and Grassland Science, National Agriculture and Food Research Organization, Ibaraki 305-0901, Japan
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MIZUTANI Eiji
National Institute of Livestock and Grassland Science, National Agriculture and Food Research Organization, Ibaraki 305-0901, Japan
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DANG-NGUYEN Thanh
Graduate School of Life and Environmental Sciences, University of Tsukuba, Ibaraki 305-8577, Japan
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Haraguchi Seiki
National Institute Of Livestock And Grassland Science (nilgs) National Agriculture And Food Research Organization (naro)
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Kaneda Masahiro
National Institute Of Livestock And Grassland Science National Agriculture And Food Research Organization
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Somfai Tamas
National Institute Of Livestock And Grassland Science
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Geshi Masaya
National Inst. Of Livestock And Grassland Sci.
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IMAI Kei
National Livestock Breeding Center, Fukushima 961-8511, Japan
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WATANABE Shinya
National Institute of Livestock and Grassland Science, National Agriculture and Food Research Organization, Ibaraki 305-0901, Japan
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AKAGI Satoshi
National Institute of Livestock and Grassland Science, National Agriculture and Food Research Organization, Ibaraki 305-0901, Japan
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INABA Yasushi
National Institute of Livestock and Grassland Science, National Agriculture and Food Research Organization, Ibaraki 305-0901, Japan
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NAGAI Takashi
National Institute of Livestock and Grassland Science, National Agriculture and Food Research Organization, Ibaraki 305-0901, Japan
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