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排卵后衰老小鼠卵母细胞中微小RNA的表达谱及功能分析

Expression profiles and function analysis of microRNAs in postovulatory aging mouse oocytes.

作者信息

Wang Tian-Yang, Zhang Jie, Zhu Jiang, Lian Hua-Yu, Yuan Hong-Jie, Gao Min, Luo Ming-Jiu, Tan Jing-He

机构信息

College of Animal Science and Veterinary Medicine, Shandong Agricultural University, Tai-an City 271018, P. R. China.

出版信息

Aging (Albany NY). 2017 Apr;9(4):1186-1201. doi: 10.18632/aging.101219.

Abstract

In this study, microRNA (miRNA) profiles in postovulatory aging mouse oocytes were analyzed by microarray screening and RT-qPCR. Hierarchical cluster analysis on the microarray data and KEGG pathway enrichment analysis on the mRNAs targeted by differentially expressed (DE) miRNAs between two adjacent egg-ages suggest that while only a mild alteration in miRNA expression occurred from 13 to 18 h, a great change took place from 18 to 24 h post hCG injection. Theoretical exploration on functions of the predicted target genes suggest that KEGG pathways enriched by 13-18 h DE miRNAs are correlated with early events of oocyte aging while pathways most enriched by 18-24 h or 24-30 h DE miRNAs are correlated with the late symptoms of aged oocytes. Experimental verification on functions of the key proteins predicted by the KEGG analysis and injection of miR-98 mimics or inhibitors further confirmed that miRNAs played stimulatory/inhibitory roles in postovulatory oocyte aging. In conclusion, marked changes in miRNA expression are associated with significant alterations in function and morphology of postovulatory aging oocytes.

摘要

在本研究中,通过微阵列筛选和逆转录定量聚合酶链反应(RT-qPCR)分析了排卵后老化小鼠卵母细胞中的微小RNA(miRNA)谱。对微阵列数据进行层次聚类分析,并对两个相邻卵龄之间差异表达(DE)miRNA靶向的mRNA进行京都基因与基因组百科全书(KEGG)通路富集分析,结果表明,虽然从13小时到18小时miRNA表达仅发生轻微变化,但在注射人绒毛膜促性腺激素(hCG)后18小时到24小时发生了巨大变化。对预测靶基因功能的理论探索表明,13 - 18小时DE miRNA富集的KEGG通路与卵母细胞老化的早期事件相关,而18 - 24小时或24 - 30小时DE miRNA最富集的通路与老化卵母细胞的晚期症状相关。对KEGG分析预测的关键蛋白功能进行实验验证,并注射miR-98模拟物或抑制剂,进一步证实了miRNA在排卵后卵母细胞老化中发挥刺激/抑制作用。总之,miRNA表达的显著变化与排卵后老化卵母细胞的功能和形态的显著改变相关。

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