营养诱导舍饲哈萨克羊非繁殖季节卵巢组织中miRNAs差异表达分析

发布时间：2018-05-29 15:17

本文选题：哈萨克羊 + 发情期　；参考：《畜牧兽医学报》2017年04期

【摘要】：旨在利用Solexa测序技术分析、挖掘营养诱导非繁殖季节绵羊发情期和乏情期卵巢组织micro RNAs(miRNA),为进一步研究特定miRNA参与非繁殖季节绵羊发情的分子调控机制提供理论基础。以不同营养水平饲喂的哈萨克绵羊作为研究对象,在非繁殖季节前后分别屠宰已鉴定的3只发情(EN)和乏情(AN)母羊,分别采集卵巢(O)组织,构建两个小片段RNA文库(OEN和OAN),进行高通量测序和生物信息学分析,实时荧光定量PCR(qRT-PCR)验证差异表达的miRNAs在哈萨克羊卵巢中表达水平。结果显示,两文库分别获得8 665 978(发情期)和9 071 102条(乏情期)clean reads,且绝大多数小RNA序列长度为21~23nt。两文库相比较,发现有9个已知和104个未知miRNAs表达差异显著(P0.01)。利用qRT-PCR随机选择的6个显著差异表达的miRNAs进行验证,其表达水平和测序分析结果一致。KEGG通路分析表明,这些差异miRNAs的靶基因主要参与了甘氨酸、丝氨酸和苏氨酸代谢,氨基糖和核苷酸糖代谢,精氨酸和鸟氨酸代谢及甘油磷脂代谢等营养调控通路。结合靶基因预测及通路富集分析,推测差异表达的miRNAs是通过营养代谢通路对绵羊的非季节性发情起重要作用。
[Abstract]:The aim of this study was to explore the micro RN AsmiRNAs in oestrus and oestrous stage of ovaries induced by nutrition in order to provide a theoretical basis for further study on the molecular regulation mechanism of specific miRNA in estrus of non-reproductive season sheep. Three identified estrous en and anestrous ANN ewes were slaughtered before and after non-breeding season in Kazak sheep fed with different nutritional levels. Two small RNA libraries were constructed for high throughput sequencing and bioinformatics analysis. The differential expression of miRNAs in Kazak ovaries was verified by real-time quantitative PCR qRT-PCRR. The results showed that 8 665.978 (estrus) and 9 071,102 (estrous) were obtained from the two libraries, and the vast majority of small RNA sequences were 21 ~ (23) nt. There were 9 known and 104 unknown miRNAs expression differences between the two libraries. Six differentially expressed miRNAs were randomly selected by qRT-PCR. The expression level was consistent with the results of sequencing analysis. KEGG pathway analysis showed that the target genes of these differentially expressed miRNAs were mainly involved in the metabolism of glycine, serine and threonine. Amino sugar and nucleotide metabolism, arginine and ornithine metabolism, glycerol phospholipid metabolism and other nutritional regulation pathways. Combined with target gene prediction and pathway enrichment analysis, it was speculated that differentially expressed miRNAs may play an important role in non-seasonal estrus of sheep through nutrient metabolic pathway.
【作者单位】：石河子大学动物科技学院;石河子大学生命科学学院;
【基金】：国家自然科学基金项目(31260537)
【分类号】：S826

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