Smith Craig S, Berg Debra K, Berg Martin, Pfeffer Peter L
AgResearch, Ruakura Campus, Hamilton, New Zealand.
Cell Reprogram. 2010 Dec;12(6):699-707. doi: 10.1089/cell.2010.0040. Epub 2010 Oct 25.
Somatic cell nuclear transfer (NT)-specific effects on postblastocyst early cattle embryogenesis were investigated by comparison to in vitro-produced (IVP) embryos grown under identical conditions to embryonic days (E) 14 and 15. Recipient effects were excluded by transferring mixed batches of NT and IVP embryos into each cow. Embryo recovery rates, proportions with an epiblast and embryo, as well as epiblast dimensions did not differ between NT and IVP embryos. A developmental expression profile was determined for nine trophoblast markers, two inner cell mass (ICM)/epiblast markers, and E-cadherin at nine time points between E7 and E26, providing a molecular gene signature assay for developmental progression. Gene expression levels for these genes (Cdx2, Elf5, Mash2, Ifn-tau, Furin, Kunitz1, Pag11, Gata3, Oct4 and Ifitm3) were equal in NT and IVP embryos of equivalent length. Furthermore, the average residual deviation of all 10 genes did not differ significantly suggesting an overall "normality" in gene expression of E14/15 NT embryos. The absence of NT-specific defects during the second, highly selective, week of cattle embryogenesis is interpreted as supportive for the view that NT-associated defects are predominantly of an epigenetic nature.
通过与在相同条件下培养至胚胎第14天和15天的体外生产(IVP)胚胎进行比较,研究了体细胞核移植(NT)对牛胚泡后早期胚胎发育的特异性影响。通过将混合批次的NT和IVP胚胎移植到每头母牛体内,排除了受体效应。NT胚胎和IVP胚胎在胚胎回收率、具有上胚层和胚胎的比例以及上胚层尺寸方面没有差异。在胚胎第7天至第26天的九个时间点,测定了九种滋养层标志物、两种内细胞团(ICM)/上胚层标志物和E-钙黏蛋白的发育表达谱,提供了一种用于发育进程的分子基因特征分析方法。在长度相当的NT胚胎和IVP胚胎中,这些基因(Cdx2、Elf5、Mash2、Ifn-tau、Furin、Kunitz1、Pag11、Gata3、Oct4和Ifitm3)的基因表达水平相等。此外,所有10个基因的平均残余偏差没有显著差异,这表明E14/15 NT胚胎的基因表达总体上“正常”。在牛胚胎发育的第二个高度选择性的星期内没有NT特异性缺陷,这被解释为支持以下观点:与NT相关的缺陷主要是表观遗传性质的。