Department of Animal Science, College of Agriculture, Yanbian University, Jilin, Yanji, 133000, China.
Institute of Animal Biotechnology, Jilin Academy of Agricultural Sciences, Jilin, Gongzhuling, 136100, China.
Theriogenology. 2022 Jul 15;187:119-126. doi: 10.1016/j.theriogenology.2022.04.025. Epub 2022 Apr 26.
RNA-seq technology can be used for the detection of miRNA transcripts in tissues and cells at specific periods and under specific treatment conditions, which can easily and effectively screen out differential transcripts. The purpose of this study was to compare miRNA expression in porcine cumulus cells before and after oocyte maturation, and to investigate the mechanism whereby cumulus cells may influence oocyte maturation. To that aim, cumulus cells surrounding GV- and MII-stage oocytes were isolated, and their differences in miRNA expression were examined using miRNA-seq. 143 differentially expressed miRNAs were identified, among which miR-101 was selected and further verified by qPCR. Moreover, miR-101 was found to target HAS2 through target gene prediction, luciferase-based co-transfection and cumulus cells transfection. Transfection of COCs with miR-101 mimics or inhibitor revealed that the miR-101 could inhibit oocyte IVM by regulating the expanding of CCSs, but had no effect on the embryo development competence. These findings demonstrated that miR-101 regulated oocyte maturation in vitro via targeting HAS2 in porcine cumulus cells.
RNA-seq 技术可用于检测特定时期和特定处理条件下组织和细胞中的 miRNA 转录本，可轻松有效地筛选出差异转录本。本研究旨在比较卵母细胞成熟前后猪卵丘细胞中的 miRNA 表达，探讨卵丘细胞影响卵母细胞成熟的机制。为此，分离了围绕 GV 和 MII 期卵母细胞的卵丘细胞，并通过 miRNA-seq 检测它们的 miRNA 表达差异。鉴定出 143 个差异表达的 miRNA，其中选择 miR-101 并通过 qPCR 进一步验证。此外，通过靶基因预测、基于荧光素酶的共转染和卵丘细胞转染发现，miR-101 可以通过靶向 HAS2 来抑制 COC 的体外 IVM，但对胚胎发育能力没有影响。这些发现表明，miR-101 通过在猪卵丘细胞中靶向 HAS2 来调节卵母细胞的成熟。
