Laboratory of Developmental Biology, Institute of Animal Physiology and Genetics of the Czech Academy of Sciences, Liběchov, Czech Republic.
Department of Animal Production, Faculty of Agriculture, Cairo University, Giza, Egypt.
Mol Reprod Dev. 2019 Apr;86(4):426-439. doi: 10.1002/mrd.23121. Epub 2019 Feb 13.
Oocyte developmental competence is acquired during folliculogenesis and regulated by complex molecular mechanisms. Several molecules are involved in these mechanisms, including microRNAs (miRNAs) that are essential for oocyte-specific processes throughout the development. The objective of this study was to identify the expression profile of miRNAs in porcine oocytes derived from follicles of different sizes using RNA deep sequencing. Oocytes were aspirated from large (LO; 3-6 mm) or small (SO; 1.5-1.9 mm) follicles and tested for developmental competence and chromatin configurations. Small RNA libraries were constructed from both groups and then sequenced in an Illumina NextSeq. 500. Oocytes from the LO group exhibited higher developmental competence and different chromatin configuration compared with oocytes from the SO group. In total, 167 and 162 known miRNAs were detected in the LO and SO groups, respectively. MiR-205, miR-16, miR-148a-3p, and miR-125b were among the top 10 highly expressed miRNAs in both groups. Eight miRNAs were differentially expressed (DE) between both groups. Target gene prediction and pathway analysis revealed 46 pathways that were enriched with miRNA-target genes. The oocyte meiosis pathway and signaling pathways including FoxO, PI3K-Akt, and cAMP were predictably targeted by DE miRNAs. These results give more insights into the potential role of miRNAs in regulating the oocyte development.
卵母细胞的发育能力是在卵泡发生过程中获得的,并受复杂的分子机制调控。有几种分子参与这些机制,包括 microRNAs(miRNAs),它们在卵母细胞发育过程中的特异性过程中是必不可少的。本研究的目的是使用 RNA 深度测序鉴定来自不同大小卵泡的猪卵母细胞中 miRNAs 的表达谱。从大卵泡(LO;3-6mm)或小卵泡(SO;1.5-1.9mm)中抽吸卵母细胞,并测试其发育能力和染色质构型。从小卵泡和大卵泡两组中构建小 RNA 文库,然后在 Illumina NextSeq 500 上进行测序。与 SO 组的卵母细胞相比,LO 组的卵母细胞表现出更高的发育能力和不同的染色质构型。在 LO 组和 SO 组中分别检测到 167 个和 162 个已知的 miRNAs。miR-205、miR-16、miR-148a-3p 和 miR-125b 是两组中表达最高的前 10 个 miRNAs 之一。有 8 个 miRNAs 在两组之间存在差异表达(DE)。靶基因预测和通路分析显示,有 46 个通路富含 miRNA 靶基因。卵母细胞减数分裂途径和信号通路,包括 FoxO、PI3K-Akt 和 cAMP,被 DE miRNAs 靶向。这些结果更深入地了解了 miRNAs 在调节卵母细胞发育中的潜在作用。