牛体外受精胚胎抗氧化相关的长链非编码RNA表达谱

doi:10.11843/j.issn.0366-6964.2017.07.007

摘要/Abstract

摘要：

旨在探究培养液中添加3 mmol·L^-1谷胱甘肽（Glutathione，GSH）对牛体外受精胚胎中长链非编码RNA（Long non-coding RNA，lncRNA）表达谱的影响。收集8~16-细胞期和桑椹期的胚胎，构建文库后测序，筛选差异lncRNAs，共表达分析后，获得的差异基因进行GO功能注释和KEGG通路分析；同时对lncRNA进行了顺式作用靶基因的预测及其功能分析；通过荧光定量PCR对8个lncRNAs表达水平进行检测，验证测序结果的准确性。测序共得到4 273个lncRNAs，通过对其表达量、长度和外显子个数分析表明，测序数据的可靠性较好。筛选共得到59个差异lncRNAs （其中23个在对照组（不添加GSH）上调，而在处理组（添加3 mmol·L^-1GSH）下调；36个在对照组下调，而在处理组上调），在对照组和处理组中，59个lncRNAs分别与754和2 782个蛋白编码基因共表达，这些基因分别参与47和48个功能分类，且主要富集于5和13条信号通路。对23和36个差异lncRNAs临近的145和154个蛋白编码基因进行GO注释，结果表明，这些基因分别参与35和43个功能分类。14个lncRNAs共表达的差异基因参与谷胱甘肽代谢通路。综上表明，在培养液中添加GSH能导致胚胎lncRNA表达谱的变化。

关键词: 谷胱甘肽, 长链非编码RNA, 转录组测序, 共表达分析, 靶基因

Abstract:

To reveal the effects of GSH supplemented in culture medium on lncRNA profiling of bovine embryos, the IVF embryos at 8-16-cell and morula stages were collected for high-throughput RNA sequencing (RNA-seq). Then co-expression analysis and cis-targeted genes prediction of differentially expressed lncRNAs (DElncRNAs), Gene ontology and KEGG pathway enrichment analysis of differentially expressed genes (DEGs) were performed. Additionally, qRT-PCR of 8 DElncRNAs was used to confirm the accuracy and validity of RNA-seq data. A total of 4 273 lncRNAs were obtained, and the qRT-PCR results confirmed the expression pattern of DElncRNAs from RNA-seq. Among those lncRNAs, 59 were differentially expressed. Among them, 23 DElncRNAs were up-regulated in control embryos while down-regulated in treated embryos, 36 DElncRNAs were down-regulated in control embryos while up-regulated in treated embryos. In control group(no GSH), 59 DElncRNAs were co-expressed with 754 DEGs which were involved in 47 GO terms and enriched in 5 pathways. In treated group, the co-expressed DEGs were 2 782 involving in 48 terms and participating in 13 pathways. Furthermore, 145 neighbor genes of 23 DElncRNAs and 154 neighbor genes of 36 DElncRNAs were participated in 35 and 43 GO terms, respectively. DEGs co-expressed with 14 lncRNAs were enriched in GSH metabolism pathways. Conclusively, GSH treatment could change the lncRNA profiling and improve the development of bovine IVF embryos.

Key words: glutathione, lncRNA, RNA-seq, co-expression analysis, target gene

中图分类号:

S823.2

郭芹芹, 李凤, 孙尉峻, 郝海生, 刘岩, 赵学明, 庞云渭, 朱化彬, 杜卫华. 牛体外受精胚胎抗氧化相关的长链非编码RNA表达谱[J]. 畜牧兽医学报, 2017, 48(7): 1229-1240.

GUO Qin-qin, LI Feng, SUN Wei-jun, HAO Hai-sheng, LIU Yan, ZHAO Xue-ming, PANG Yun-wei, ZHU Hua-bin, DU Wei-hua. Long Non-Coding RNA Profiling in Bovine Embryos Treated with Glutathione during in vitro Culture[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(7): 1229-1240.

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