2、6 和 12 月龄小尾寒羊卵巢中 miRNA 和 mRNA 表达谱的综合分析显示，oar-miR-432 下调 RPS6KA1 的表达。

Integrated analysis of miRNA and mRNA expression profiles in 2-, 6-, and 12-month-old Small Tail Han Sheep ovaries reveals that oar-miR-432 downregulates RPS6KA1 expression.

机构信息

College of Animal Science and Technology, Jinlin Agricultural University, Changchun, Jilin, China.

出版信息

Gene. 2019 Aug 20;710:76-90. doi: 10.1016/j.gene.2019.02.095. Epub 2019 Mar 18.

DOI:10.1016/j.gene.2019.02.095

Abstract

Small Tail Han Sheep are an excellent local sheep breed in China, and their outstanding reproductive performance is one of their very important biological characteristics. Clarifying the ovary development process of these ewes should provide a theoretical basis for improving their reproductive efficiency. In this study, we identified the differentially expressed (DE) microRNAs (miRNAs) in 2-, 6-, and 12-month-old small-tail Han sheep ovaries by constructing and analyzing the miRNA expression profiles. These findings clarify the molecular mechanisms regulating the excellent reproductive performance of small-tail Han ewes. We used RNA-Seq technology and bioinformatic to analyze these profiles. Eleven, 13, and 19 DE miRNAs were identified in the 2- vs 6-, 6- vs 12-, and 2- vs 12-month-old ovaries, respectively. In total, 54, 37, and 198 predicted target genes of these DE miRNAs were identified in these three groups, respectively. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses showed that in the 2- vs 6-month-old ovaries, the target genes of DE known sheep miRNAs were involved in 102 GO terms and seven signaling pathways; in the 6- vs 12-month-old ovaries, the target genes of DE known sheep miRNAs were involved in 52 GO terms and three signaling pathways; and in the 2- vs 12-month-old ovaries, the target genes of DE known sheep miRNAs were involved in 88 GO terms and six signaling pathways. Three miRNA-target regulatory networks were constructed based on these DE miRNA-target interactions. Nine miRNAs were selected to confirm to the accuracy of the miRNA sequencing data with qRT-PCR. The site at which oar-miR-432 binds RPS6KA1 was determined with a dual-luciferase system. This is the first integrated analysis the expression profiles of miRNAs and their targets during ovarian development in small-tail Han sheep. These data clarify the molecular regulatory mechanisms underlying sheep ovarian development and identify biomarkers that influence the reproductive performance of small-tail Han ewes.

摘要

小尾寒羊是中国优秀的绵羊品种之一，其突出的繁殖性能是其非常重要的生物学特征之一。阐明小尾寒羊卵巢的发育过程，可为提高其繁殖效率提供理论依据。本研究通过构建和分析 miRNA 表达谱，鉴定了 2 月龄、6 月龄和 12 月龄小尾寒羊卵巢中的差异表达（DE）microRNAs（miRNAs）。这些发现阐明了调控小尾寒羊优良繁殖性能的分子机制。我们使用 RNA-Seq 技术和生物信息学方法对这些图谱进行了分析。在 2 月龄与 6 月龄、6 月龄与 12 月龄、2 月龄与 12 月龄卵巢中分别鉴定到 11、13 和 19 个 DE miRNAs。这三组分别鉴定到这些 DE miRNAs 的 54、37 和 198 个预测靶基因。GO 和 KEGG 分析表明，在 2 月龄与 6 月龄卵巢中，DE 已知绵羊 miRNAs 的靶基因涉及 102 个 GO 术语和 7 个信号通路；在 6 月龄与 12 月龄卵巢中，DE 已知绵羊 miRNAs 的靶基因涉及 52 个 GO 术语和 3 个信号通路；在 2 月龄与 12 月龄卵巢中，DE 已知绵羊 miRNAs 的靶基因涉及 88 个 GO 术语和 6 个信号通路。基于这些 DE miRNA-靶相互作用构建了三个 miRNA-靶调控网络。用 qRT-PCR 对 9 个 miRNA 进行验证，以确定 miRNA 测序数据的准确性。利用双荧光素酶系统确定了 oar-miR-432 与 RPS6KA1 结合的位点。这是首次对小尾寒羊卵巢发育过程中 miRNA 及其靶基因的表达谱进行的综合分析。这些数据阐明了绵羊卵巢发育的分子调控机制，并鉴定了影响小尾寒羊繁殖性能的生物标志物。