Subsets of cloned mouse embryos and their non-random relationship to development and nuclear reprogramming
Subsets of cloned mouse embryos and their non-random relationship to development and nuclear reprogramming
复制标题
DOI：
10.1016/j.mod.2007.09.007
复制
发表时间：
2008-01-01
影响因子：
2.6
通讯作者：
Boiani, Michele
中科院分区：
文献类型：
--
作者：
Cavaleri, Fatima M.;Balbach, Sebastian T.;Boiani, Michele
在卵母细胞介导的核重新编程中，一个重要的问题是供体核的基因表达是随机变化还是遵循某种模式。由于克隆胚胎是非常异质的，在植入前发育过程中经常停滞，因此随机情况被普遍接受。在本研究中，我们通过识别结构亚集来解决卵丘细胞来源的小鼠克隆的异质性，并分析了它们与供体核重新编程的关系。我们利用Oct4启动子驱动的GFP转基因的活细胞成像来解析克隆的和ICSI受精的桑椹胚群体，并根据不同的GFP表达将它们分成三个亚集。在功能上，克隆的但不是ICSI桑拿子的亚群形成囊胚和ES细胞，与Oct4-GFP的表达成正比。多能性相关基因Oct4、Nanog和Sox2的转录水平差异很小。不同亚群中选择的mRNAs水平的微小差异表明，从桑椹胚阶段开始，重新编程的过程是一致的，而不是随机的。由于这些微小的差异与发育能力的显著差异相对应，我们认为克隆的发育潜力与基因表达水平的关系不同于受精胚胎。(C)2007爱思唯尔爱尔兰有限公司。保留所有权利。
An important question in oocyte-mediated nuclear reprogramming is whether gene expression of the donor nucleus changes randomly or follows a pattern. Since cloned embryos are very heterogenous and arrest frequently during preimplantation development, a random scenario is generally accepted. In the present study, we resolve the heterogeneity of cumulus cell-derived mouse clones by recognizing structured subsets, and we analyze their relationship to reprogramming of donor nuclei. We utilize live cell imaging of the Oct4 promoter-driven GFP transgene to resolve the populations of cloned and ICSI-fertilized morulae, and we sort them both into three subsets based on different GFP expression. Functionally, subsets of cloned but not ICSI morulae form blastocysts and ES cells proportional to Oct4-GFP expression. Regulatively, the subsets of cloned morulae are characterized by small differences of transcript level for the pluripotency-associated genes Oct4, Nanog and Sox2. Small differences of the level of select mRNAs across subsets suggest a uniform rather than random course of reprogramming from the morula stage on. Since these small differences correspond with substantial differences in developmental competence, we propose that developmental potential of clones relates to levels of gene expression in a different way than fertilized embryos. (c) 2007 Elsevier Ireland Ltd. All rights reserved.