Molecular Endocrinology, Functional Genomics and Systems Biology Laboratory, Animal Biochemistry Division, National Dairy Research Institute, Karnal, Haryana, India.
J Cell Biochem. 2018 Nov;119(10):7934-7943. doi: 10.1002/jcb.26508. Epub 2018 Jun 28.
Ovarian folliculogenesis, ovulation, and luteinization are an important prerequisite for fertility performance in mammals. Spatial and temporal key factors and proteins for their regulation are well known. Recent advancement in the field of molecular biology led to the discovery of another class of gene regulators, microRNA (miRNA). Previous studies on profiling of miRNA in buffalo ovaries revealed that miRNA-210 (miR-210) is differently expressed in follicular-luteal transition. Therefore, the present study was planned to ascertain the role of miR-210 in buffalo granulosa cells. Cultured granulosa cells were transfected with miR-210 mimic. Effect of overexpression of miR-210 was analyzed on granulosa cell marker genes (CYP19A1 and PCNA) which were significantly downregulated (P < 0.05). Further, target genes of miR-210 were screened using Target Scan software v7.1 and a list of 37 genes with cumulative weight context score (CWCS) > 0.5 was sorted followed by their functional annotation and network analyses using PANTHER and STRING software. Bioinformatics analyses identified HRas gene as a potential hub gene of miR-210 targeted genes. HRas has been shown to be involved in diverse biological pathways regulating ovarian functions. An expression analysis of HRas was further validated both in vitro and in vivo. EFNA3 (EFHRIN-A3), another identified target of miR-210 known to be involved in angiogenesis, was also downregulated in miR-210 transfected granulosa cells. In conclusion, the present study demonstrated that miR-210 can regulate granulosa cell function at preovulatory stage through HRas and EFNA3. Further studies are needed to find the mechanism how miR-210 regulates the granulosa cells function through these targets.
哺乳动物的卵巢卵泡发生、排卵和黄体化是生育能力表现的重要前提。其调节的时空关键因素和蛋白已众所周知。分子生物学领域的最新进展导致了另一类基因调控因子，microRNA（miRNA）的发现。以前在水牛卵巢中miRNA 谱分析的研究表明，miR-210（miR-210）在卵泡-黄体转变中表达不同。因此，本研究旨在确定 miR-210 在水牛颗粒细胞中的作用。用 miR-210 模拟物转染培养的颗粒细胞。分析过表达 miR-210 对颗粒细胞标记基因（CYP19A1 和 PCNA）的影响，这些基因显著下调（P<0.05）。此外，使用 Target Scan 软件 v7.1 筛选 miR-210 的靶基因，根据累积权重上下文评分（CWCS）>0.5 对 37 个基因进行排序，然后使用 PANTHER 和 STRING 软件进行功能注释和网络分析。生物信息学分析确定 HRas 基因是 miR-210 靶基因的潜在枢纽基因。HRas 已被证明参与调节卵巢功能的多种生物学途径。进一步在体外和体内验证了 HRas 的表达分析。EFNA3（EFHRIN-A3），另一个已知参与血管生成的 miR-210 靶基因，也在 miR-210 转染的颗粒细胞中下调。总之，本研究表明，miR-210 可以通过 HRas 和 EFNA3 调节排卵前阶段的颗粒细胞功能。需要进一步的研究来发现 miR-210 通过这些靶基因调节颗粒细胞功能的机制。
