犏牛雄性不育相关lncRNA的鉴定与分析

doi:10.11843/j.issn.0366-6964.2019.03.010

摘要/Abstract

摘要：

旨在分析成年健康牦牛和犏牛（3~4岁）睾丸组织中长链非编码RNA（long non-coding RNA，lncRNA）的表达谱，从而探究lncRNA与犏牛雄性不育机制的关联性，为解释犏牛雄性不育的机理提供理论依据。采集成年（3~4岁）健康牦牛、犏牛各3头的睾丸组织，构建cDNA文库后进行高通量测序，筛选出差异的lncRNA，通过对差异显著的lncRNA与mRNA位置关系以及结合能的判定预测顺式作用（cis）和反式作用（tran）的靶基因，并进行GO、KEGG功能分析。结果表明，共得到牦牛和犏牛候选lncRNA转录本分别为20 363和24 133个，两个文库筛选出6 178个显著差异lncRNA，其中有2 470个在犏牛睾丸组织与牦牛比较上调，3 708个下调。筛选得到差异显著lncRNA的候选靶基因2 676个，这些基因参与58个功能分类，共涉及306条通路；其中主要富集的通路包括轴突指导、内吞、Hedgehog等信号通路。综上表明，部分lncRNA介导的靶基因PTGDS、IGF2、MEST、GLIS3、NOTCH2、HOXA10、HOXA11与犏牛雄性不育相关，lncRNA在犏牛的生殖发育中对其雄性不育具有重要的调控作用。利用RNA-Seq技术筛选出成年牦牛和犏牛睾丸组织lncRNA，并进行差异分析，为探讨犏牛雄性不育的机制提供更完善的转录组数据。

Abstract:

The objective of this study was to explore the link between lncRNAs and the mechanism of male infertility of cattle-yak by analyzing the gene expression profile about lncRNA in testis tissue of adult healthy yak and cattle-yak (3-4 years old). This study provided the base for further interpreting the mechanism of male infertility of cattle-yak.Testis tissues of 3 adult healthy yaks and 3 adult healthy cattle-yaks were collected to construct the cDNA libraries. And, the structure and function of genes were analyzed by using high-throughput sequencing and differentially expressed lncRNAs (DElncRNAs) were screened between yak and cattle-yak. Then the cis-targeted and tran-targeted genes of DElncRNAs were predicted by exploring the positional relation and binding energy between lncRNA and mRNA, and the GO and KEGG pathway enrichment analysis was also made. The results displayed that 20 363 and 24 133 candidate lncRNA transcripts of yak and cattle-yak were respectively obtained, 6 178 DElncRNAs of which were selected.Compared with yak, there were 2 470 up-regulated and 3 708 down-regulated lncRNAs in cattle-yak. Meanwhile 2 676 candidate target genes of DElncRNAs were obtained. The analytical results indicated that these genes participated in 58 functional classifications and participated in 306 signal pathways. And, they were mainly involved in such signal pathways as axon guidance, endocytosis and hedgehog. In conclusion, these findings inferred that some lncRNA-mediated target genes (PTGDS, IGF2, MEST, GLIS3, NTOCH2, HOXA10, HOXA11) were associated with male infertility of cattle-yak, and lncRNA may played an important role in the reproductive development of cattle-yak. Moreover, the gap analysis of lncRNA was made by using RNA-Seq technology,which provided more complete data for further exploring the mechanisms of male infertility of cattle-yak on transcriptome.

中图分类号:

S823.8⁺5.2

阿果约达, 熊显荣, 王艳, 杨显英, 韩杰, 王斌, 李键. 犏牛雄性不育相关lncRNA的鉴定与分析[J]. 畜牧兽医学报, 2019, 50(3): 551-561.

A GUO Yueda, XIONG Xianrong, WANG Yan, YANG Xianying, HAN Jie, WANG Bin, LI Jian. Identification and Analysis of Long Non-coding RNA Associated with Cattle-yak Male Infertility[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(3): 551-561.

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