Zielak-Steciwko A E, Browne J A, McGettigan P A, Gajewska M, Dzięcioł M, Szulc T, Evans A C O
Institute of Animal Breeding, Wrocław University of Environmental and Life Sciences, Wrocław, Poland;
School of Agriculture and Food Science, University College Dublin, Belfield, Dublin, Ireland;
Physiol Genomics. 2014 Oct 1;46(19):735-45. doi: 10.1152/physiolgenomics.00036.2014. Epub 2014 Aug 5.
Development of ovarian follicles is controlled at the molecular level by several gene products whose precise expression leads to regression or ovulation of follicles. MicroRNAs (miRNAs) are small noncoding RNAs that regulate gene expression through sequence-specific base pairing with target messenger RNAs (mRNAs) causing translation repression or mRNA degradation. The aim of this study was to identify miRNAs expressed in theca and/or granulosa layers and their putative target genes/pathways that are involved in bovine ovarian follicle development. By using miRCURY microarray (Exiqon) we identified 14 and 49 differentially expressed miRNAs (P < 0.01) between dominant and subordinate follicles in theca and granulosa cells, respectively. The expression levels of four selected miRNAs were confirmed by qRT-PCR. To identify target prediction and pathways of differentially expressed miRNAs we used Union of Genes option in DIANA miRPath v.2.0 software. The predicted targets for these miRNAs were enriched for pathways involving oocyte meiosis, Wnt, TGF-beta, ErbB, insulin, P13K-Akt, and MAPK signaling pathways. This study identified differentially expressed miRNAs in the theca and granulosa cells of dominant and subordinate follicles and implicates them in having important roles in regulating known molecular pathways that determine the fate of ovarian follicle development.
卵巢卵泡的发育在分子水平上受几种基因产物的控制，这些基因产物的精确表达会导致卵泡退化或排卵。微小RNA（miRNA）是一类小的非编码RNA，通过与靶信使RNA（mRNA）进行序列特异性碱基配对来调节基因表达，从而导致翻译抑制或mRNA降解。本研究的目的是鉴定在牛卵泡膜和/或颗粒层中表达的miRNA及其假定的靶基因/途径，这些基因/途径参与牛卵巢卵泡的发育。通过使用miRCURY微阵列（Exiqon），我们分别在卵泡膜细胞和颗粒细胞中鉴定出优势卵泡和从属卵泡之间有14个和49个差异表达的miRNA（P < 0.01）。通过qRT-PCR证实了四个选定miRNA的表达水平。为了鉴定差异表达miRNA的靶标预测和途径，我们在DIANA miRPath v.2.0软件中使用了基因联合选项。这些miRNA的预测靶标在涉及卵母细胞减数分裂、Wnt、TGF-β、ErbB、胰岛素、PI3K-Akt和MAPK信号通路的途径中富集。本研究鉴定了优势卵泡和从属卵泡的卵泡膜细胞和颗粒细胞中差异表达的miRNA，并表明它们在调节决定卵巢卵泡发育命运的已知分子途径中具有重要作用。
