Single-cell gene expression of the bovine blastocyst.
Single-cell gene expression of the bovine blastocyst.
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DOI：
10.1530/rep-17-0345
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发表时间：
2017-11
期刊：
影响因子：
--
通讯作者：
Hansen PJ
中科院分区：
文献类型：
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作者：
Negrón-Pérez VM;Zhang Y;Hansen PJ
胚胎中的前两个分化事件导致三种细胞类型-上胚层、滋养外胚层（TE）和下胚层。本研究的目的是鉴定牛中每种细胞类型的分子标记物，并评估每种谱系的单个细胞之间的基因表达差异。来自67个单个细胞的解离胚泡的cDNA被用来确定93个基因的转录本丰度，这些基因在其他物种中作为细胞谱系标志物或可能参与发育过程。聚类分析表明，该细胞属于两个主要群体（进化枝A和B），其中进化枝A有两个亚群，进化枝B有四个亚群。利用来自其他物种的谱系特异性标记表明，进化枝A的两个亚群分别代表上胚层和下胚层，而进化枝B的四个亚群则代表TE。在上胚层中上调的基因包括AJAP 1、DNMT 3A、FGF 4、H2 AFZ、KDM 2B、NANOG、POU 5 F1、SAV 1和SLIT 2。在下胚层中过表达的基因包括ALPL、FGFR 2、FN 1、GATA 6、GJA 1、HDAC 1、MBNL 3、PDGFRA和SOX 17，而在所有四个TE群体中过表达的基因是ACTA 2、CDX 2、CYP 11 A1、GATA 2、GATA 3、IFNT、KRT 8、RAC 1和SFN。TE的亚群对于包括原型TE标记IFNT在内的多个基因彼此不同。鉴定了牛囊胚中每种细胞类型的新标记。结果还表明TE细胞中基因表达的异质性。需要进一步的研究来确认TE细胞亚群是否代表了定型TE表型发育的不同阶段。
The first two differentiation events in the embryo result in three cell types – epiblast, trophectoderm (TE), and hypoblast. The purpose here was to identify molecular markers for each cell type in the bovine and evaluate differences in gene expression among individual cells of each lineage. The cDNA from 67 individual cells of dissociated blastocysts was used to determine transcript abundance for 93 genes implicated as cell lineage markers in other species or potentially involved in developmental processes. Clustering analysis indicated that the cells belonged to two major populations (clades A and B) with two subpopulations of clade A and four of clade B. Use of lineage-specific markers from other species indicated that the two subpopulations of clade A represented epiblast and hypoblast, respectively while the four subpopulations of clade B were TE. Among the genes upregulated in epiblast were AJAP1, DNMT3A, FGF4, H2AFZ, KDM2B, NANOG, POU5F1, SAV1 and SLIT2. Genes overexpressed in hypoblast included ALPL, FGFR2, FN1, GATA6, GJA1, HDAC1, MBNL3, PDGFRA and SOX17 while genes overexpressed in all four TE populations were ACTA2, CDX2, CYP11A1, GATA2, GATA3, IFNT, KRT8, RAC1 and SFN. The subpopulations of TE varied amongst each other for multiple genes including the prototypical TE marker IFNT. New markers for each cell type in the bovine blastocyst were identified. Results also indicate heterogeneity in gene expression among TE cells. Further studies are needed to confirm whether subpopulations of TE cells represent different stages in development of a committed TE phenotype.