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MG132 treatment during oocyte maturation improves embryonic development after somatic cell nuclear transfer and alters oocyte and embryo transcript abundance in pigs

机译:卵母细胞成熟过程中的MG132处理可改善体细胞核转移后的胚胎发育,并改变猪的卵母细胞和胚胎转录本丰度

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The objective of this study was to examine the effect of treating pig oocytes during in vitro maturation (IVM) with a proteasome inhibitor, MG132, on oocyte maturation and embryonic development. In one series of experiments, oocytes from medium-sized follicles (3-8mm in diameter) were untreated (MCO) or treated with MG132 during 0-22hr (M0-22) or 30-42hr (M3042) of IVM. There was no significant effect of MG132 on nuclear maturation or cytoplasmic maturation (as assessed by intracellular amounts of glutathione and p34cdc2 kinase activity). Blastocyst formation after parthenogenetic activation (PA) and somatic cell nuclear transfer (SCNT), however, was increased for M3042 (65.2% and 27.7% for PA and SCNT, respectively) compared to MCO (42.6% and 13.6%, respectively) and M022 (45.3% and 19.5%, respectively; P<0.05). Expression of PCNA and ERK2 was increased in M3042 for IVM oocytes while transcript abundance for POUF51, DNMT1, FGFR2, and PCNA was increased in M3042 for 4-cell SCNT embryos. When oocytes derived from small follicles (<3mm in diameter) were untreated (SCO) or treated with MG132 during 0-22hr (S0-22), 30-42hr (S30-42) of IVM, or 0-22 and 30-42hr of IVM (S0-22/30-42), expression of POU5F1, DNMT1, FGFR2, and PCNA and blastocyst formation were increased for SCNT embryos derived from S30 to 42 (16.5%) and S0-22/30-42 oocytes (20.8%) as compared to embryos from SCO (8.7%) or S022 oocytes (8.8%; P<0.05). Results demonstrate that treatment of oocytes with MG132 during the later stage of IVM improves embryonic development and alters gene expression in pigs. Mol. Reprod. Dev. 79:4150, 2012. (C) 2011 Wiley Periodicals, Inc.
机译:这项研究的目的是研究使用蛋白酶体抑制剂MG132在体外成熟(IVM)过程中处理猪卵母细胞对卵母细胞成熟和胚胎发育的影响。在一系列实验中,在IVM的0-22小时(M0-22)或30-42小时(M3042)期间,未处理(MCO)或用MG132处理来自中型卵泡(直径3-8mm)的卵母细胞。 MG132对核成熟或细胞质成熟没有显着影响(通过细胞内谷胱甘肽的量和p34cdc2激酶活性评估)。然而,与MCO(分别为42.6%和13.6%)和M022相比,M3042(单性生殖激活(PA)和体细胞核转移(SCNT)之后的胚泡形成增加了(PA和SCNT分别为65.2%和27.7%)。 (分别为45.3%和19.5%; P <0.05)。 IVM卵母细胞在M3042中PCNA和ERK2的表达增加,而4细胞SCNT胚胎在M3042中POUF51,DNMT1,FGFR2和PCNA的转录本丰度增加。如果在0-22小时(S0-22),30-42小时(S30-42)的IVM或0-22和30-42小时内未处理(SCO)或用MG132处理源自小卵泡的卵母细胞(直径小于3mm) IVM(S0-22 / 30-42)的变化,POU5F1,DNMT1,FGFR2和PCNA的表达以及胚泡形成对于S30至42(16.5%)和S0-22 / 30-42卵母细胞(20.8)的SCNT胚胎增加与来自SCO(8.7%)或S022卵母细胞(8.8%; P <0.05)的胚胎相比。结果表明,在IVM后期,用MG132处理卵母细胞可改善猪的胚胎发育并改变基因表达。大声笑责备。开发人员79:4150,2012.(C)2011 Wiley Periodicals,Inc.

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