解剖学报

解剖学报 ›› 2021, Vol. 52 ›› Issue (1): 124-129.doi: 10.16098/j.issn.0529-1356.2021.01.020

• 组织学胚胎学发育生物学 • 上一篇    下一篇

小鼠4倍体胚胎诱导蜕膜化反应

张斌 焦熙堯 李华 张静 王浩洋 周荣艳 李相运*   

  1. 河北农业大学动物科技学院，河北省牛羊胚胎技术创新中心，河北 保定  071000
  • 收稿日期:2019-08-16 修回日期:2019-10-30 出版日期:2021-02-06 发布日期:2021-02-06
  • 通讯作者: 李相运 E-mail:lxyun@hebau.edu.cn
  • 基金资助:
    国家重点研发计划项目

Decidualization induced by mouse tetraploid embryos

ZHANG Bin  JIAO Xi-yao  LI Hua  ZHANG Jing  WANG Hao-yang  ZHOU Rong-yan  LI Xiang-yun*   

  1. College of Animal Science and Technology, Research Center of Cattle and Sheep Embryo Engineering Technique of Hebei, Hebei Agricultural University, Hebei Baoding  071000, China
  • Received:2019-08-16 Revised:2019-10-30 Online:2021-02-06 Published:2021-02-06
  • Contact: LI Xiang-yun E-mail:lxyun@hebau.edu.cn

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摘要：

目的  探讨小鼠囊胚内细胞团（ICM）在蜕膜化过程中是否具有调控作用。  方法  采用电融合技术制备4倍体胚胎，作为内细胞团缺陷胚胎，通过输卵管胚胎移植制备4倍体胚胎诱导蜕膜组织，以正常2-细胞胚胎诱导的蜕膜组织作为对照，从着床位点和蜕膜形态对两组蜕膜进行比较分析后，采用高通量测序筛选两组中差异表达的微小RNA (miRNA)，生物信息学分析两组蜕膜中差异miRNA的靶基因，进行基因功能注释和通路富集分析。  结果  4倍体胚胎诱导的蜕膜组织与2-细胞胚胎诱导蜕膜组织在着床位点极为相似，但是蜕膜形态差异存在显著性。两组蜕膜中差异表达miRNA共有16个，与2-细胞胚胎诱导蜕膜相比，11个miRNA（miR-466f-3p、miR-302 d-3p、miR-466i-5p、miR-465c-5p、miR-302a-5p、miR-7068-3p、miR-741-3p、miR-302a-3p、miR-433-5p、miR-144-5p、miR-878-5p）在4倍体胚胎诱导蜕膜中表达上调，5个miRNA（miR-690、miR-193b-5p、miR-147-3p、novel_327、miR-363-3p）在4倍体胚胎诱导蜕膜中表达下调。生物信息学分析显示，差异miRNA功能主要集中在蛋白结合、离子束缚等功能，靶基因主要参与环磷酸鸟苷蛋白激酶G(cGMP-PKG)信号通路、雌激素信号通路、血管内皮生长因子(VEGF)信号通路等。  结论  内细胞团在蜕膜化反应过程中具有重要调控作用。

关键词: 4倍体胚胎, 蜕膜, 微小RNA, 内细胞团, 高通量测序, 小鼠

Abstract:

Objective  To investigate the role of inner cell mass (ICM) in decidualization using decidua induced by two-cell embryos or tetraploid embryos through tubal embryo transfer.   Methods  Tetraploid embryo, as the inner cell mass-deficient embryo, were produced by electrofusion of mouse 2-cell embryos. Decidua was induced by 2-cell embryos or tetraploid embryos through tubal embryo transfer. Decidua induced by 2-cell embryos was employed as a control. Morphologic and implantation site of decidua were compared between two-cell embryo-induced decidua and tetraploid embryo-induced decidua. The differentially expressed microRNA (miRNA) was screened by high-throughput sequencing. The target genes of differentially expressed miRNA in two groups were screened for Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG).   Results  Tetraploid embryo-induced decidual tissue and 2-cell embryo-induced decidual tissue were very similar at the implantation site, but there were significant differences in decidual morphology. There were 16 miRNAs differentially expressed in decidua of the two groups, of which 11 miRNAs（miR-466f-3p，miR-302 d-3p，miR-466i-5p，miR-465c-5p, miR-302a-5p, miR-7068-3p, miR-741-3p, miR-302a-3p, miR-433-5p, miR-144-5p, miR-878-5p）were up-regulated in tetraploid embryo-induced decidua and 5 miRNAs（miR-690, miR-193b-5p, miR-147-3p, novel_327, miR-363-3p）were down-regulated in tetraploid embryo-induced decidua. Bioinformatics analysis showed that the target genes played function such as protein binding and ion binding, and mainly involved in cyclic guanosine monophosphate-protein kinase G(cGMP-PKG) signaling pathway, estrogen signaling pathway, vascular endothelial growth factor (VEGF) signaling pathway and so on.   Conclusion  ICM plays an important role in decidualization.

Key words: Tetraploid, Decidua, MicroRNA, Inner cell mass, High-throughput sequencing, Mouse

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