激素处理后绵羔羊和成年母羊卵巢颗粒细胞miRNA特征分析  

Comparative miRNA Analysis of Granulosa Cells in Lamb and Ewe with Superstimulation Treatment

作　　者：林嘉鹏[1,2] 吴阳升 韩冰[1] 李晓林[1,2] 汪立芹[1] 刘明军[1] 黄俊成[1] LIN Jia-peng;WU Yang-sheng;HAN Bing;LI Xiao-lin;WANG Li-qin;LIU Ming-jun;HUANG Jun-cheng(Ministry of Agriculture,Livestock Breeding and Genetics and Breeding Key Laboratory,Xinjiang Academy of Animal Sciences Biotech nology Research Center,Xinjiang Urumqi 830000,China;College of Animal Science and Technology,Shihezi University,Xinjiang Shi hezi 832003,China)

机构地区：[1]农业部草食家畜遗传育种与繁殖重点开放实验室新疆维吾尔自治区畜牧科学院生物技术研究所,新疆乌鲁木齐830000 [2]石河子大学动物科技学院,新疆石河子832003

出　　处：《西南农业学报》2018年第11期2423-2430,共8页Southwest China Journal of Agricultural Sciences

基　　金：新疆维吾尔自治区自然科学基金(2016D01B003)

摘　　要：【目的】通过small RNA高通量测序技术,筛选绵羔羊和成年母羊差异表达miRNA,进一步挖掘和鉴定与卵泡发育相关的相关基因,揭示其性状差异的信号通路及分子调控网络。【方法】以绵羔羊和成年母羊颗粒细胞为研究对象,应用高通量测序技术进行small RNA文库构建,获得miRNA表达谱,鉴定差异表达miRNA,进而对这些差异表达miRNA及其靶基因进行功能注释、聚类分析及通路富集等。【结果】从2个样本miRNA库鉴定出238个已知miRNAs和79个新miRNAs。共获得9个差异miRNA,与成年羊相比,羔羊有4个miRNA上调,5个下调。其中,oar-miR-106a上调倍数最大(1. 85),oar-miR-10a下调最为显著(-2. 20)。对差异miRNAs进行GO富集和KEGG通路富集分析表明,新陈代谢途径富集的基因最多。利用qRT-PCR对5个随机挑选的miRNA进行验证,结果与测序数据一致,说明测序结果可靠。【结论】miRNAs可能在激素处理后对绵羊生殖生理中起着重要的作用,包括卵泡和卵母细胞的发育,颗粒细胞的增殖等。[ Objective]The present paper was conducted to screen differentially expressed miRNA from lambs and adult ewes through modem molecular biology and high-throughput sequencing technology, and further dig out and identify related genes related to follicular develop- ment, and reveal signaling pathways and molecular regulatory networks of their differences in traits. [ Method ] With lamb and ewe granulosa cells as the research object, the high-throughput sequencing （RNA-Seq） technology for RNA libral7 construction was applied to reveal the genetic differences between the lamb and adult sheep from the transcriptome level, the identification of differentially expressed miRNA, and the differences of expression of MI RNA and its target gene functional annotation and clustering analysis and pathway enrichment etc were studied. [ Result] More than 200 miRNAs were identified from two miRNA libraries, and 1/3 of them were identified as new miRNAs. Of the 9 differences in the conunon identification of miRNA, the relative ewes, the lamb had 4 miRNA up-regulated and 5 down-regulation. Among them, the up regulation of oar-miR-106a was the most significant （1.85） , and the oar-miR-10a downregulation was the most significant （ - 2.20）. The GO enrichment and KEGG pathway analysis of all the target genes of all miRNAs in two libraries showed that the abundance of PI3K/Akt signaling pathway was the highest. Real time fluorescence quantitative PCR comirmed 5 randomly selected miRNA, and the re- suits were consistent with the sequencing data. [ Conclusion] miRNAs may play an important role in the reproductive physiology of sheep after superovulation, including the development of follicle and oocyte and the proliferation of granulosa cells.

关 键 词：绵羔羊 成年母羊 MICRORNA 卵泡 

分 类 号：S826[农业科学—畜牧学]