Functional genomics of 5- to 8-cell stage human embryos by blastomere single-cell cDNA analysis.

Functional genomics of 5- to 8-cell stage human embryos by blastomere single-cell cDNA analysis.
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DOI:
10.1371/journal.pone.0013615
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发表时间:
2010-10-26
期刊:
影响因子:
3.7
通讯作者:
Simón C
Simón C
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Galán A;Montaner D;Póo ME;Valbuena D;Ruiz V;Aguilar C;Dopazo J;Simón C

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人类胚胎发育过程中的卵裂球命运和胚胎基因组激活(EGA)是具有高度科学和临床意义的未解决领域。采用高效的单细胞 cDNA 扩增方案对来自 5 至 8 细胞人类胚胎的 49 个卵裂球进行了研究,为高密度微阵列分析提供了模板。对之前描述的内细胞团 (ICM) (n = 120)、干性 (n = 190) 和滋养外胚层 (TE) (n = 45) 特征标记进行了分析,并建立了 46 个基因的管家模式。来自 5 至 8 细胞阶段胚胎的所有人类卵裂球均显示出与 ICM 标记(例如 DDX3、FOXD3、LEFTY1、MYC、NANOG、POU5F1)、干性(例如 POU5F1、DNMT3B、GABRB3、SOX2、ZFP42、TERT)和 TE 标记(例如 GATA6、 EOMES、CDX2、LHCGR)。还研究了 5-6 细胞和 8 细胞阶段胚胎的 EGA 谱,并与囊胚阶段进行比较。确认了已知基因(n = 92),例如耗尽的母体转录本(例如,CCNA1、CCNB1、DPPA2)和胚胎特异性激活(例如,POU5F1、CDH1、DPPA4)以及新基因。总之,对 5 至 8 细胞阶段人类胚胎的整体单细胞 cDNA 扩增微阵列分析表明,卵裂球命运并不取决于 ICM 或 TE。最后,提出了人类胚胎发生中新的 EGA 特征。
Blastomere fate and embryonic genome activation (EGA) during human embryonic development are unsolved areas of high scientific and clinical interest. Forty-nine blastomeres from 5- to 8-cell human embryos have been investigated following an efficient single-cell cDNA amplification protocol to provide a template for high-density microarray analysis. The previously described markers, characteristic of Inner Cell Mass (ICM) (n = 120), stemness (n = 190) and Trophectoderm (TE) (n = 45), were analyzed, and a housekeeping pattern of 46 genes was established. All the human blastomeres from the 5- to 8-cell stage embryo displayed a common gene expression pattern corresponding to ICM markers (e.g., DDX3, FOXD3, LEFTY1, MYC, NANOG, POU5F1), stemness (e.g., POU5F1, DNMT3B, GABRB3, SOX2, ZFP42, TERT), and TE markers (e.g., GATA6, EOMES, CDX2, LHCGR). The EGA profile was also investigated between the 5-6- and 8-cell stage embryos, and compared to the blastocyst stage. Known genes (n = 92) such as depleted maternal transcripts (e.g., CCNA1, CCNB1, DPPA2) and embryo-specific activation (e.g., POU5F1, CDH1, DPPA4), as well as novel genes, were confirmed. In summary, the global single-cell cDNA amplification microarray analysis of the 5- to 8-cell stage human embryos reveals that blastomere fate is not committed to ICM or TE. Finally, new EGA features in human embryogenesis are presented.
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发表时间: 2004-07-13
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