植入前囊胚滋养外胚层基因空间表达

发布时间：2018-03-20 11:49

本文选题：囊胚　切入点：极滋养层细胞　出处：《北京大学学报(医学版)》2017年06期 　论文类型：期刊论文

【摘要】：目的:研究人胚胎植入前囊胚滋养外胚层细胞基因的空间表达。方法:辅助生殖来源人受精后第6天Gardner评分5AA的囊胚,显微镜下微吸管机械分离滋养外胚层细胞,对囊胚滋养外胚层极滋养层细胞和壁滋养层细胞采用单细胞测序检测,选取经t检验计算所得P0.05且基因表达差异≥2倍的差异表达基因,采用生物信息学软件对筛选的差异基因进行无监督层次聚类分析和基因本体(gene ontology,GO)功能分类分析,对差异表达基因进行注释和生物功能富集,并用全基因组对差异基因功能进一步注释。结果:对2枚受精后第6天的5AA囊胚滋养外胚层8个极滋养层细胞和8个壁滋养层细胞测序,发现极滋养层细胞306个基因上调,壁滋养层细胞75个基因上调。无监督聚类分析结果显示外胚层细胞分成极滋养层细胞和壁滋养层细胞两群,属于两种不同类型和生物功能的细胞群。差异基因功能注释显示,极滋养层细胞上调基因GO生物学功能主要是转录、能量代谢、蛋白合成、转运、氧化应激、离子转运、蛋白合成及运输、细胞周期调节、肌动蛋白增长等,主要参与泛素介导的蛋白水解、氧化磷酸化、Wnt信号通路、雌雄激素代谢等信号通路;壁滋养层细胞上调基因GO生物学功能主要是蛋白分解代谢、细胞周期停滞、细胞凋亡、激活丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)信号通路、碳水化合物运输、突触负调节、细胞生长、钙通道激活、正向B细胞分化、T细胞凋亡等,主要参与B细胞受体、T细胞受体、白细胞跨内皮移植、血管内皮生长因子(vascular endothelial growth factor,VEGF)表达、间隙连接、促性腺激素释放激素(gonadotropin-releasing hormone,GnRH)分泌、细胞凋亡等信号通路。结论:从空间维度揭示囊胚极/壁滋养层细胞的基因表达,显示胚胎植入前囊胚极/壁滋养外胚层相互协调,精细调节囊胚孵化和胚胎植入过程,进一步数据分析有望寻找到调控胚胎植入的内源性特异分子。
[Abstract]:Objective: to study the spatial expression of human blastocyst trophodermal cell gene in preimplantation human blastocyst. Methods: the blastocyst with Gardner score of 5AA on the 6th day after assisted reproduction was used to isolate the trophoblast ectodermal cells by micropipette mechanically. Single cell sequencing was used to detect trophoblast cells and parietal trophoblast cells of blastocyst trophoderm. Bioinformatics software was used for unsupervised hierarchical cluster analysis and functional classification analysis of gene ontologyGO. the differentially expressed genes were annotated and biofunctional enrichment was carried out. Results: on the 6th day after fertilization, the 5AA blastocysts were sequenced from 8 extremely trophoblast cells and 8 parietal trophoblast cells, and 306 genes were up-regulated in the very trophoblast cells. The results of unsupervised cluster analysis showed that the ectodermal cells were divided into two groups of extremely trophoblastic cells and parietal trophoblastic cells, which belonged to two different types and biological function groups. The up-regulation of go gene in trophoblast cells mainly includes transcription, energy metabolism, protein synthesis, transport, oxidative stress, ion transport, protein synthesis and transport, cell cycle regulation, actin growth and so on. Ubiquitin mediated protein hydrolysis, oxidative phosphorylation of Wnt signaling pathway, androgen metabolism, etc. The up-regulated gene go biological function of trophoblastic cells is mainly protein catabolism, cell cycle arrest, cell apoptosis, etc. Activation of mitogen-activated protein kinases (MAPK) signaling pathway, carbohydrate transport, synaptic negative regulation, cell growth, calcium channel activation, positive differentiation of T cells into B cells, apoptosis of T cells, etc. The expression of vascular endothelial growth factor (VEGF), gap junction, gonadotropin-releasing hormone gonadotropin-releasing hormone (GnRH), vascular endothelial growth factor (VEGF), vascular endothelial growth factor (VEGF) and gonadotropin-releasing hormone (GnRH) were observed in leukocyte transendothelial transplantation. Conclusion: the gene expression of blastocyst / parietal trophoblast cells is revealed from the spatial dimension, which indicates that the blastocyst pole / wall trophoblast ectoderm is coordinated with each other, and the hatching and implantation process of blastocyst are carefully regulated. Further data analysis is expected to find endogenous specific molecules regulating embryo implantation.
【作者单位】：北京积水潭医院妇产科;
【基金】：国家自然科学基金(81070493)资助~~
【分类号】：R714.8

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