Abstract
Mastitis is a common disease in dairy cows that is mostly caused by E. coli, and it brings massive losses to the dairy industry. N6-Methyladenosine (m6A), a methylation at the N6 position of RNA adenine, is a type of modification strongly associated with many diseases. However, the role of m6A in mastitis has not been investigated. In this study, we used MeRIP-seq to sequence the RNA of bovine mammary epithelial cells treated with inactivated E. coli for 24 h. In this in vitro infection model, there were 16,691 m6A peaks within 7066 mRNA transcripts in the Con group and 10,029 peaks within 4891 transcripts in the E. coli group. Compared with the Con group, 474 mRNAs were hypermethylated and 2101 mRNAs were hypomethylated in the E. coli group. Biological function analyses revealed differential m6A-modified genes mainly enriched in the MAPK, NF-κB, and TGF-β signaling pathways. In order to explore the relationship between m6A and mRNA expression, combined MeRIP-seq and mRNA-seq analyses revealed 212 genes with concomitant changes in the mRNA expression and m6A modification. This study is the first to present a map of RNA m6A modification in mastitis treated with E. coli, providing a basis for future research.
Highlights
Mastitis is one of the most severe diseases in dairy farming, and its diagnosis and treatment present many challenges
MAC-T cells were treated with heat-inactivated E. coli with an multiplicity of infection (MOI) of 10:1 for 24 h
The results show that the expression levels of proinflammatory factors IL-1β, IL-6, and TNF-α were significantly increased in the MAC-T cells treated with inactivated E. coli (p < 0.05)
Summary
Mastitis is one of the most severe diseases in dairy farming, and its diagnosis and treatment present many challenges. Mammary epithelial cells (MECs), as the first barrier of nonspecific immunity of mastitis, play a crucial role in mastitis following infection by pathogenic microorganisms. These cells secrete a variety of inflammatory factors and chemokines [3] that regulate cell apoptosis, the Toll-like receptor (TLR) pathway, the mitogen-activated protein kinase (MAPK) pathway, and other signaling pathways [4]. We determined potential m6 A modification in the inflammation of bovine mammary epithelial cells treated with inactivated E. coli using high-throughput sequencing (MeRIP-seq). We investigated the relationship between m6 A modification and mRNA transcription These findings lay a foundation for further exploration of m6 A RNA modification in mastitis treated with E. coli
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