发情周期不同阶段绵羊卵巢lncRNA的鉴定和功能分析

旨在分析不同繁殖周期绵羊卵巢组织中长链非编码RNA (long non-coding RNA，lncRNA)的表达谱，了解lncRNA表达及其调控机理，为绵羊繁育研究提供理论依据。本研究以湖羊为研究对象，选取年龄在1.5～2.5岁的黄体期和卵泡期母羊各3只，通过高通量测序筛选出卵巢组织中的lncRNA，运用生物信息学分析对差异表达的lncRNA进行靶基因预测，通过GO和KEGG富集分析找出与绵羊繁殖相关的通路。结果显示，本研究共获得1 379个差异表达的lncRNAs，其中1 158个表达上调，221个表达下调。GO和KEGG富集分析表明，差异表达的lncRNAs及其靶基因主要参与卵泡发育、排卵周期过程、钙离子信号通路、卵母细胞减数分裂、催产素信号通路、MAPK信号通路、甲状腺激素合成通路、雌激素信号通路。关键的lncRNAs可能通过调控参与这些信号通路和生物学过程的相关基因来调控生殖。其中，LNC_011239、LNC_012847、LNC_003902、LNC_003906、LNC_003907等靶向的MAPK1、ADCY1、ADCY5、PPP3CA和CDC23可能发挥关键的调控作用。qRT-PCR验证证明，随机选取的5个差异lncRNAs定量结果与测序结果基本一致。本研究利用RNA-Seq技术筛选出黄体期和卵泡期卵巢组织中的lncRNAs，并进行差异分析，为揭示绵羊繁殖能力的分子机制提供依据。

Identification and Functional Analysis of lncRNA in Ovaries of Sheep at Different Stages of Estrus Cycle

This study aimed to analyze the expression profiles of long non-coding RNA (lncRNA) in sheep ovarian tissues at different reproductive cycles, understand the lncRNA expression and its regulatory mechanism, and provide a theoretical basis for sheep breeding research. In this study, Hu sheep were selected as the research object, and 3 ewes aged from 1.5 to 2.5 years old in luteal phase and follicular phase were selected, respectively. lncRNA in ovarian tissues were screened by high-throughput sequencing. Bioinformatics analysis was used to predict the target genes of differentially expressed lncRNAs. GO and KEGG enrichment analysis were used to identify pathways related to sheep reproduction. The results showed that a total of 1 379 differentially expressed lncRNAs were obtained in this study, of which 1 158 were up-regulated and 221 down-regulated. GO and KEGG enrichment analysis showed that the differentially expressed lncRNAs and their target genes mainly involved in ovarian follicle development, ovulation cycle process, calcium signaling pathway, oocyte meiosis, oxytocin signaling pathway, MAPK signaling pathway, thyroid hormone synthesis signaling pathway, and estrogen signaling pathway. Key lncRNAs might regulate reproduction by regulating genes involved in these signaling pathways and biological processes. Specifically, MAPK1, ADCY1, ADCY5, PPP3CA and CDC23, which are targeted by LNC_011239, LNC_012847, LNC_003902, LNC_003906, LNC_003907 and others, might play key regulatory roles. The results of qRT-PCR showed that the expression levels of 5 randomly selected lncRNAs were consistent with the sequencing results. In this study, RNA-Seq technology was used to screen lncRNAs in luteal and follicular ovarian tissues, and the differences were analyzed to provide the basis for revealing the molecular mechanism of sheep reproductive capacity.