小尾寒羊间情期和发情期microRNAs差异表达分析

doi:10.11843/j.issn.0366-6964.2016.07.004

摘要/Abstract

摘要：

本研究通过构建和分析小尾寒羊间情期和发情期卵巢组织microRNA（miRNA）表达谱，筛选两个时期差异表达的microRNAs，为研究microRNA调控小尾寒羊繁殖过程提供相应的理论基础。利用活体手术法在发情期（第1天）和间情期（第13天）分别采集一侧卵巢。从卵巢中提取总RNA，利用illumina Hiseq2000测序平台获取RNA数据，对表达谱和差异表达microRNAs进行生物信息学分析。实时荧光定量PCR（qRT-PCR）验证差异表达的microRNAs在小尾寒羊卵巢中表达水平。结果，成功地构建出小尾寒羊间情期和发情期microRNA的表达谱，oar-miR-99a和oar-miR-143分别是间情期和发情期表达量最高的microRNA，并筛选出在两个时期间3个显著差异表达的microRNAs，分别为oar-miR-200a、oar-miR-200b和oar-miR-200c。利用qRT-PCR对随机选择的2个显著差异表达的microRNAs进行验证，其表达水平和RNA-Seq分析结果一致。microRNA表达谱为后续的绵羊卵巢microRNA研究提供更详尽的信息。结合靶基因预测及通路富集分析，推测差异表达的microRNAs是通过代谢和免疫途径调控卵巢周期性活动。

Abstract:

To explore the theoretical basis of the microRNAs in regulating the reproductive process in Small Tail Han sheep，the ovarian microRNAs profiles were constructed and analyzed in estrus and diestrus.One ovary was collected with surgery method at estrus (the 1st day) and diestrus (the 13th day) from the same sheep，respectively.The RNA sequencing data were obtained with illumina Hiseq2000 sequencing platform.The expression profiles and differential expression of microRNAs were analyzed by bioinformatics method.The real-time quantitative PCR (qRT-PCR) was used for validating the differentially expressed microRNAs between the 2 periods in ovary.The microRNAs expression profiles of Small Tailed Han sheep was constructed in the period of estrus and diestrus in ovaries.The highest expression of microRNA was observed for oar-miR-99a and oar-miR-143 in the period of diestrus and estrus，respectively.Three microRNAs，oar-miR-200a，oar-miR-200b and oar-miR-200c，were significantly differentially expressed between estrus and diestrus.The expression level of 2 randomly selected microRNAs by qRT-PCR was the same as the result by RNA-seq data.These microRNAs expression profiles will be used to provide informations for further studying the microRNAs in sheep ovary.The differentially expressed microRNAs may regulate the ovarian periodic activity through the metabolism and immunity pathway by the analysis of predicted target genes and enriched pathways.

中图分类号:

S826
S813.3

段新崇，魏彦辉，李阳，李相运，周荣艳，锡建中. 小尾寒羊间情期和发情期microRNAs差异表达分析[J]. 畜牧兽医学报, 2016, 47(7): 1324-1332.

DUAN Xin-chong，WEI Yan-hui，LI Yang，LI Xiang-yun，ZHOU Rong-yan，XI Jian-zhong. Analyzing the Differential Expression of microRNAs in Estrus and Diestrus of Small Tailed Han Sheep[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(7): 1324-1332.

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