Laboratory of Animal Breeding and Reproduction, Graduate School of Agriculture, Hokkaido University, Sapporo, Japan.
Biol Reprod. 2013 Jun 27;88(6):158. doi: 10.1095/biolreprod.113.108993. Print 2013 Jun.
Mice and cattle use distinct pathways for the first cell segregation into inner cell mass (ICM) and trophectoderm (TE) lineages at the blastocyst stage. However, limited knowledge is available regarding the reliable transcriptional networks that orchestrate the complex developmental processes at this stage in nonrodent species. In order to elucidate the site-dominant transcriptomic properties of bovine blastocysts, we separated cell samples into the ICM and TE using both mechanical and chemical methods and performed in silico prescreening for candidate genes that were site-dominantly expressed in bovine blastocysts. We further performed quantitative real-time PCR and in situ hybridization using the site-specific cell samples. As a result, we identified seven ICM-dominant genes and five TE-dominant genes not found in earlier studies. Our findings provide novel insights into the mechanism of cell-fate specification in the pre-implantation bovine embryo.
在囊胚阶段，老鼠和牛使用不同的途径将细胞第一次分离成内细胞团（ICM）和滋养外胚层（TE）谱系。然而，关于在非啮齿动物物种中协调这一复杂发育过程的可靠转录网络，我们知之甚少。为了阐明牛囊胚中主导位点的转录组特性，我们使用机械和化学方法将细胞样本分离为 ICM 和 TE，并进行了候选基因的计算机预筛选，这些候选基因在牛囊胚中表现出主导位点的表达。我们进一步使用特定部位的细胞样本进行了定量实时 PCR 和原位杂交。结果，我们鉴定了七个 ICM 优势基因和五个早期研究中未发现的 TE 优势基因。我们的研究结果为着床前牛胚胎细胞命运特化的机制提供了新的见解。
