Reproduction Lab and IVF Unit, Department of Obstetrics and Gynecology, Sheba Medical Center, 52662, Tel Hashomer, Affiliated with the Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel.
Department of Molecular Biology, Ariel University, Ariel, Israel.
Sci Rep. 2018 Oct 23;8(1):15605. doi: 10.1038/s41598-018-33807-y.
Ovarian follicular development and ovulation are complex and tightly regulated processes that involve regulation by microRNAs (miRNAs). We previously identified differentially expressed mRNAs between human cumulus granulosa cells (CGCs) from immature early antral follicles (germinal vesicle - GV) and mature preovulatory follicles (metaphase II - M2). In this study, we performed an integrated analysis of the transcriptome and miRNome in CGCs obtained from the GV cumulus-oocyte complex (COC) obtained from IVM and M2 COC obtained from IVF. A total of 43 differentially expressed miRNAs were identified. Using Ingenuity IPA analysis, we identified 7288 potential miRNA-regulated target genes. Two hundred thirty-four of these target genes were also found in our previously generated ovulatory gene library while exhibiting anti-correlated expression to the identified miRNAs. IPA pathway analysis suggested that miR-21 and FOXM1 cooperatively inhibit CDC25A, TOP2A and PRC1. We identified a mechanism for the temporary inhibition of VEGF during ovulation by TGFB1, miR-16-5p and miR-34a-5p. The linkage bioinformatics analysis between the libraries of the coding genes from our preliminary study with the newly generated library of regulatory miRNAs provides us a comprehensive, integrated overview of the miRNA-mRNA co-regulatory networks that may play a key role in controlling post-transcriptomic regulation of the ovulatory process.
卵巢卵泡发育和排卵是一个复杂且受到严密调控的过程,其中涉及 microRNAs(miRNAs)的调控。我们之前已经鉴定出人类初级窦卵泡(生发泡期-GV)和成熟排卵前卵泡(M2 期)的卵丘颗粒细胞(CGC)之间差异表达的 mRNAs。在这项研究中,我们对从 IVM 获得的 GV 卵丘-卵母细胞复合物(COC)和从 IVF 获得的 M2 COC 的 CGC 进行了转录组和 miRNome 的综合分析。共鉴定出 43 个差异表达的 miRNA。使用 Ingenuity IPA 分析,我们鉴定出了 7288 个潜在的 miRNA 调控靶基因。其中 234 个靶基因也存在于我们之前生成的排卵基因文库中,并且与鉴定出的 miRNA 表达呈负相关。IPA 通路分析表明,miR-21 和 FOXM1 协同抑制 CDC25A、TOP2A 和 PRC1。我们发现 TGFB1、miR-16-5p 和 miR-34a-5p 通过暂时抑制 VEGF 来促进排卵。我们对初步研究中的编码基因文库与新生成的调控 miRNA 文库之间的链接生物信息学分析,为我们提供了一个全面的综合概述,即 miRNA-mRNA 共同调控网络,可能在控制排卵过程中转录后调控中发挥关键作用。