RNA-seq转录组测序分析不同季节对槟榔江水牛精液品质的影响

doi:10.11843/j.issn.0366-6964.2016.07.009

摘要/Abstract

摘要：

本研究旨在利用RNA-seq技术对不同季节槟榔江水牛全血mRNA进行分析，找出不同季节精液品质候选基因。选取5头生长状况相近的3~5岁槟榔江水牛，进行春、夏、秋、冬4个季节的精液品质（鲜精活力、精子畸形率、射精量、精子密度）检测。根据精液品质，选取差异极显著的春（BL）、夏（SBL）两个季节的水牛样品进行RNA-seq转录组测序分析。颈静脉采集全血，抽提mRNA并组内混合制样，利用RNA-seq技术分析不同季节槟榔江水牛全血mRNA差异表达。根据差异倍数>1.5，P<0.05的标准进行筛选，得到差异基因，再利用DAVID 6.7软件对差异基因进行GO和KEGG分析，通过RT-PCR验证基因芯片结果的准确性。结果表明：差异表达基因84个，其中上调基因56个，下调基因28个；GO分析发现，这些差异基因与应激反应、免疫反应有关；KEGG分析表明，差异基因显著富集到肿瘤坏死因子（TNF）和T细胞受体信号通路。其中，神经营养因子受体类似物死亡结构域（NRADD，又叫热休克蛋白因子1 (HSF1)）、原癌基因FOS和JUN，与精液品质相关性强，可作为进一步研究的靶基因。

关键词: RNA-seq, 槟榔江水牛, 差异表达基因, 精液品质

Abstract:

To determine the key candidate genes related to semen quality in different seasons，the mRNA profiling of Chinese Betelnut-jiang buffalo were analyzed by RNA-seq.Based on similarities in growth status，5 Betelnut-jiang buffalo bulls were selected and their semen quality were tested (including ejaculate volume，fresh sperm motility，sperm density and abnormality rate).According to the semen quality，the samples of summer (treated as SBL group) and spring (as BL group) were selected for RNA-seq.We collected the whole blood in jugular vein of Betelnut-jiang buffalo bulls，extracted the mRNAs and mixed them within group.Then，the mRNA profiling was used to measure the mRNAs in Betelnut-jiang buffalo bulls.Gene expression levels with >1.5 fold change and P value < 0.05 between the BL and SBL groups were further analyzed.Furthermore，GO and KEGG were used to analyze the genes enrichment.Finally，RT-PCR was used to confirm the RNA-seq result.The results indicated 84 differentially expressed genes.Among these，56 genes were up-regulated and 28 genes were down-regulated in SBL group.The GO search revealed that these differential expression genes were mainly related to stress reaction and immune response.And the KEGG selected the TNF signaling pathway and T cell receptor signaling pathway.Neurotrophin receptor alike death domain（NRADD，also named heat shock factor protein 1(HSF1)），localisation of FOS (FOS) and jun proto-oncogene (JUN)，were strongly associated with semen quality in different seasons which could be used as target genes for further study.

Key words: RNA-seq, Betelnut-jiang buffalo, differently expressed genes, semen quality

中图分类号:

S823.8+3

耿红红，张敬旸，李莲，王根林. RNA-seq转录组测序分析不同季节对槟榔江水牛精液品质的影响[J]. 畜牧兽医学报, 2016, 47(7): 1373-1380.

GENG Hong-hong，ZHANG Jing-yang，LI Lian，WANG Gen-lin. Semen Quality Analysis of Betelnut-jiang Buffalo in Different Seasons by RNA-seq[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(7): 1373-1380.

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