Genome-wide methylation and transcriptome of blood neutrophils reveal the roles of DNA methylation in affecting transcription of protein-coding genes and miRNAs in E. coli-infected mastitis cows
文献类型: 外文期刊
作者: Ju, Zhihua 1 ; Jiang, Qiang 1 ; Wang, Jinpeng 1 ; Wang, Xiuge 1 ; Yang, Chunhong 1 ; Sun, Yan 1 ; Zhang, Yaran 1 ; Wang, Changfa 1 ; Gao, Yaping 1 ; Wei, Xiaochao 1 ; Hou, Minghai 1 ; Huang, Jinming 1 ;
作者机构: 1.Shandong Acad Agr Sci, Dairy Cattle Res Ctr, Jinan 250131, Shandong, Peoples R China
2.Engn Ctr Anim Breeding & Reprod, Jinan 250100, Shandong, Peoples R China
关键词: Cows; E; coli mastitis; Neutrophils; DNA methylation; Expression pattern; miRNA; Alternative splicing; Immune responses
期刊名称:BMC GENOMICS ( 影响因子:3.969; 五年影响因子:4.478 )
ISSN: 1471-2164
年卷期: 2020 年 21 卷 1 期
页码:
收录情况: SCI
摘要: Background Neutrophils are the first effectors of inflammatory response triggered by mastitis infection, and are important defense cells against pathogenic Escherichia coli (E. coli). DNA methylation, as a critical epigenetic mechanism for regulating gene function, is involved in bovine mastitis. Results In this study, we sequenced the blood neutrophils of healthy and E. coli-infected mastitic half-sib cows for the overall DNA methylation levels using transcriptome sequencing and reduced representation bisulfite sequencing. The methylation levels in the mastitis cows (MCs) were decreased compared with healthy cows (HCs). A total of 494 differentially methylated regions were identified, among which 61 were up-methylated and 433 were down-methylated (MCs vs. HCs). The expression levels of 1094 differentially expressed genes were up-regulated, and 245 genes were down-regulated. Twenty-nine genes were found in methylation and transcription data, among which seven genes' promoter methylation levels were negatively correlated with expression levels, and 11 genes were differentially methylated in the exon regions. The bisulfite sequencing PCR and quantitative real-time PCR validation results demonstrated that the promoter methylation of CITED2 and SLC40A1 genes affected differential expression. The methylation of LGR4 exon 5 regulated its own alternative splicing. The promoter methylation of bta-miR-15a has an indirect effect on the expression of its target gene CD163. The CITED2, SLC40A1, and LGR4 genes can be used as candidates for E. coli-induced mastitis resistance. Conclusions This study explored the roles of DNA methylation in affecting transcription of protein-coding genes and miRNAs in E. coli-induced mastitis, thereby helping explain the function of DNA methylation in the pathogenesis of mastitis and provided new target genes and epigenetic markers for mastitis resistance breeding in dairy cattle.
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