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首页> 外文期刊>The Japanese Journal of Veterinary Research >Effect of activation treatments of recipient oocytes on subsequent development of bovine nuclear transfer embryos
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Effect of activation treatments of recipient oocytes on subsequent development of bovine nuclear transfer embryos

机译：受体卵母细胞活化处理对牛核移植胚胎后续发育的影响

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摘要

Effects of recipient oocyte activation methods on the development of nuclear transfer (NT) embryos were investigated. In Exp. 1, cell-cycle phase of serum-starved bovine cumulus cells was examined by flow cytometry. Majority(95.5 percent) of medium-sized (16-20 mu m) cells that made up 56 percent of total cells was at the G_0/G_1 phase. NT embryos were constructed by electric fusion with the medium-sized serum-starved cumulus cells and bovine oocytes of 3 different preparations: enucleated oocytes treated with calcium ionophore A 23187 for 5 min and cycloheximide for 5 hr (A 23187/CHX), those treated with ethanol for 7 min and cycloheximide for 2 hr (ethanol/CHX) and those without treatment. In Exp. 2 and 3, developmentalcompetence of NT embryos constructed with A 23187/CHX- and ethanol/CHX-treated oocytes was compared to that of NT embryos constructed with non-treated oocytes, respectively. Further, nuclear behavior in 3 different NT embryos was examined in Exp. 4. Within 1 hr after fusion, majority of the NT embryos constructed with non-treated oocytes showed condensed chromosome. Three hours after fusion, about 50 percent of NT embryos constructed with non-treated or ethanol/CHX-treated oocytes showed a single pronucleus-like structure. NT embryos constructed with ethanol/CHX-treated oocytes showed similar rates of fusion, cleavage and blastocyst formation to those of the non-treated oocytes. In contrast, NT embryos constructed with A 23187/CHX-treated oocytes did not show any pronucleus-like structure and showed lower cleavage rate and no development to blastocysts. The results indicate that ethanol/CHX-treated oocytes could support development of somatic cell NT embryos to the blastocyst stage at a similar rate to that of non-treated oocytes.
机译：研究了受体卵母细胞激活方法对核移植（NT）胚胎发育的影响。在实验中参照图1，通过流式细胞术检查血清饥饿的牛卵丘细胞的细胞周期期。占总细胞56％的中型细胞（16-20μm）的大多数（95.5％）处于G_0 / G_1阶段。 NT胚胎通过与3种不同制剂的中型血清饥饿的卵丘细胞和牛卵母细胞电融合而构建：用钙离子载体A 23187处理5分钟的去核卵母细胞和环己酰亚胺5小时（A 23187 / CHX）处理的卵母细胞用乙醇处理7分钟，然后用环己酰亚胺处理2小时（乙醇/ CHX），然后不进行处理。在实验中参照图2和图3，分别比较了用A 23187 / CHX和乙醇/ CHX处理的卵母细胞构建的NT胚与未处理的卵母细胞构建的NT胚的发育能力。此外，在Exp。中检查了3个不同NT胚胎中的核行为。 4.融合后1小时内，用未经处理的卵母细胞构建的大部分NT胚胎显示染色体浓缩。融合三小时后，用未经处理的或经乙醇/ CHX处理的卵母细胞构建的NT胚胎中约有50％呈现出单个原核样结构。用乙醇/ CHX处理的卵母细胞构建的NT胚胎显示出与未处理的卵母细胞相似的融合，卵裂和胚泡形成速率。相比之下，用A 23187 / CHX处理的卵母细胞构建的NT胚胎没有显示任何前核样结构，并且显示出较低的裂解率，也没有发育为胚泡。结果表明，乙醇/ CHX处理的卵母细胞可以以与未处理卵母细胞相似的速率支持体细胞NT胚胎发育到胚泡期。

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