A mRNA landscape of bovine embryos after standard and MAPK-inhibited culture conditions: a comparative analysis.
A mRNA landscape of bovine embryos after standard and MAPK-inhibited culture conditions: a comparative analysis.
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DOI：
10.1186/s12864-015-1448-x
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发表时间：
2015-04-10
期刊：
影响因子：
4.4
通讯作者：
Roelen BA
中科院分区：
文献类型：
--
作者：
Brinkhof B;van Tol HT;Groot Koerkamp MJ;Riemers FM;IJzer SG;Mashayekhi K;Haagsman HP;Roelen BA
参与多能性的基因和信号通路已经在小鼠和人类植入前胚胎和胚胎干细胞（ES）中得到了广泛的研究。从包括牛在内的其他物种中产生胚胎干细胞系的失败尝试表明，其他基因和途径参与维持这些物种的多能性。为了研究哪些基因参与了牛的多能性，我们使用微阵列分析从桑葚胚、囊胚、滋养外胚层和内细胞团（ICM）样品中生成了表达谱。由于抑制MAPK可以增加内细胞团中NANOG/GATA6的比例，因此我们在MAPK抑制剂存在的情况下培养囊胚，分析基因在内细胞团中的表达变化。桑葚胚与囊胚间差异表达3774个基因，囊胚上调基因差异最大。基因本体论（GO）分析表明，脂质代谢过程是囊胚中表达水平较高的基因最丰富的术语。桑葚胚中表达水平较高的基因在RNA加工GO期富集。在比较ICM和TE的497个差异表达基因中，NANOG、SOX2和POU5F1在ICM中的表达增加，证实了它们在多能性中的进化保存作用。与分化或命运决定有关的几个基因在ICM中也以更高的水平表达。ICM中高水平表达的基因在RNA剪接和基因表达调控方面富集。抑制MAPK后，NANOG的表达升高，而SOX2和POU5F1的表达没有明显升高。MAPK通路中的其他基因，包括DUSP4和SPRY4，或受MAPK抑制影响的IFNT的表达下调。从微阵列研究中获得的数据为牛胚胎发育过程中的基因表达提供了进一步的见解。它们在多能性细胞中表现出表达谱，表明多能性、上皮细胞样状态。在MAPK活性抑制剂存在的情况下，ICM细胞不能作为干细胞培养，这表明仅抑制MAPK不足以维持牛细胞的多能性。本文的在线版本（doi:10.1186/s12864-015-1448-x）包含补充材料，可供授权用户使用。
Genes and signalling pathways involved in pluripotency have been studied extensively in mouse and human pre-implantation embryos and embryonic stem (ES) cells. The unsuccessful attempts to generate ES cell lines from other species including cattle suggests that other genes and pathways are involved in maintaining pluripotency in these species. To investigate which genes are involved in bovine pluripotency, expression profiles were generated from morula, blastocyst, trophectoderm and inner cell mass (ICM) samples using microarray analysis. As MAPK inhibition can increase the NANOG/GATA6 ratio in the inner cell mass, additionally blastocysts were cultured in the presence of a MAPK inhibitor and changes in gene expression in the inner cell mass were analysed. Between morula and blastocyst 3,774 genes were differentially expressed and the largest differences were found in blastocyst up-regulated genes. Gene ontology (GO) analysis shows lipid metabolic process as the term most enriched with genes expressed at higher levels in blastocysts. Genes with higher expression levels in morulae were enriched in the RNA processing GO term. Of the 497 differentially expressed genes comparing ICM and TE, the expression of NANOG, SOX2 and POU5F1 was increased in the ICM confirming their evolutionary preserved role in pluripotency. Several genes implicated to be involved in differentiation or fate determination were also expressed at higher levels in the ICM. Genes expressed at higher levels in the ICM were enriched in the RNA splicing and regulation of gene expression GO term. Although NANOG expression was elevated upon MAPK inhibition, SOX2 and POU5F1 expression showed little increase. Expression of other genes in the MAPK pathway including DUSP4 and SPRY4, or influenced by MAPK inhibition such as IFNT, was down-regulated. The data obtained from the microarray studies provide further insight in gene expression during bovine embryonic development. They show an expression profile in pluripotent cells that indicates a pluripotent, epiblast-like state. The inability to culture ICM cells as stem cells in the presence of an inhibitor of MAPK activity together with the reported data indicates that MAPK inhibition alone is not sufficient to maintain a pluripotent character in bovine cells. The online version of this article (doi:10.1186/s12864-015-1448-x) contains supplementary material, which is available to authorized users.
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影响因子：
11.8
作者：
Chazaud, Claire;Yamanaka, Yojiro;Rossant, Janet
通讯作者：
Rossant, Janet
DOI：
10.1073/pnas.0402597101
发表时间：
2004-07-13
影响因子：
11.1
作者：
Hamatani, T;Daikoku, T;Dey, SK
通讯作者：
Dey, SK
影响因子：
3.7
作者：
Kim JH;Jee MK;Lee SY;Han TH;Kim BS;Kang KS;Kang SK
通讯作者：
Kang SK
影响因子：
3.7
作者：
Khan DR;Dubé D;Gall L;Peynot N;Ruffini S;Laffont L;Le Bourhis D;Degrelle S;Jouneau A;Duranthon V
通讯作者：
Duranthon V
agriGO：农业界的 GO 分析工具包
影响因子：
14.9
作者：
Du Z;Zhou X;Ling Y;Zhang Z;Su Z
通讯作者：
Su Z