Tissue organization alters gene expression in equine induced trophectoderm cells
Tissue organization alters gene expression in equine induced trophectoderm cells
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DOI：
10.1016/j.ygcen.2017.01.030
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发表时间：
2017-06-01
影响因子：
2.7
通讯作者：
Johnson, Sally E.
中科院分区：
文献类型：
--
作者：
Reinholt, Brad M.;Bradley, Jennifer S.;Johnson, Sally E.
在哺乳动物囊胚形成的早期，形态和基因表达发生快速变化。囊胚成功保留和妊娠的关键是一个功能性的滋养外胚层（TE），它为发育中的胚胎提供旁分泌因子和激素。在马诱导的滋养细胞（iTr）细胞中检测了TE构象变化对基因表达的贡献。将马iTr细胞培养成单层或悬浮成球。球体是中空的，结构上让人联想到马的囊胚。从iTr单层和球中分离总RNA，并进行RNA测序分析。研究人员分别分析了平均3220万个和3100万个球形和单层的对齐reads。44个基因为单层所特有，45个基因仅在球中表达。构象不影响天然TE的核心基因网络CDX2、POU5F1、TEAD4、ETS2、ELF3、GATA2或TFAP2A的表达。生物信息学分析用于鉴定在响应组织形状变化时差异表达的基因类别。在iTr球和单层中，大多数差异表达基因与细胞、发育和代谢过程的结合活性相关。固有的蛋白质：蛋白质相互作用，在iTr细胞中发现了几个受体-配体家族，它们富集了编码p13激酶和MAPK信号中间体的基因。我们的研究结果为早期营养外胚层结构变化背后的配体启动激酶信号通路提供了证据。(c) 2017 Elsevier Inc.版权所有。
Rapid morphological and gene expression changes occur during the early formation of a mammalian blastocyst. Critical to successful retention of the blastocyst and pregnancy is a functional trophectoderm (TE) that supplies the developing embryo with paracrine factors and hormones. The contribution of TE conformational changes to gene expression was examined in equine induced trophoblast (iTr) cells. Equine iTr cells were cultured as monolayers or in suspension to form spheres. The spheres are hollow and structurally reminiscent of native equine blastocysts. Total RNA was isolated from iTr monolayers and spheres and analyzed by RNA sequencing. An average of 32.2 and 31 million aligned reads were analyzed for the spheres and monolayers, respectively. Forty-four genes were unique to monolayers and 45 genes were expressed only in spheres. Conformation did not affect expression of CDX2, POU5F1, TEAD4, ETS2, ELF3, GATA2 or TFAP2A, the core gene network of native TE. Bioinformatic analysis was used to identify classes of genes differentially expressed in response to changes in tissue shape. In both iTr spheres and monolayers, the majority of the differentially expressed genes were associated with binding activity in cellular, developmental and metabolic processes. Inherent to protein:protein interactions, several receptor-ligand families were identified in iTr cells with enrichment of genes coding for P13-kinase and MAPK signaling intermediates. Our results provide evidence for ligand initiated kinase signaling pathways that underlie early trophectoderm structural changes. (c) 2017 Elsevier Inc. All rights reserved.