Project description:We report the results of MIRA-Seq based high-throughput profiling of the bovine dermal fibroblast methylome from two different breeds of cattle (n=4/breed) to determine the breed-dependent differences in methylation.
Project description:Bovine rotavirus (BRV) and bovine coronavirus (BCV) infect intestinal villous epithelium in young cattle. A surgical model was adapted for neonatal calves in which a region of the jejunum was isolated from the digestive tract but lymph drainage, enervation and blood flow were maintained. Replicate sections of intestine (loops) were infected with either BRV or BCV and adjacent segments were injected with phosphate-buffered saline. Tissues were collected 18 hours post-infection. Four animals were used for BRV infection studies, and three animals were used for BCV infection studies. Microarray analyses provided a global evaluation of host gene expression patterns following BRV and BCV infection and changes in gene expression were validated by qRT-PCR analyses.
Project description:Bovine Rotavirus (BRV) causes massive economic losses in the livestock industry worldwide. MA-104 cell line has become a convenient tool for discover BRV–host interactions. Notwithstanding, the role of miRNAs in MA-104 cells during BRV infection is still ambiguous. In our study, we performed Illumina RNA sequencing analysis of miRNA libraries of BRV-infected and mock-infected MA-104 cells (at different time points 0 hpi (just after adsorption time (90 min)), 6 hpi, 12 hpi, 24 hpi, 36 hpi and 48 hpi), and the total clean reads 74701041 and 74184124 were obtained from BRV-infected and uninfected cells, respectively. From these findings, 579 categorized as known miRNAs and 144 as novel miRNAs, 160 differentially expressed (DE) miRNAs in BRV-infected in comparison to uninfected MA-104 cells were successfully investigated, 95 were upregulated and 65 exhibited downregulation. The target mRNAs of DE miRNAs were expected by bioinformatics. The functional annotation of the target genes by GO and KEGG suggested that they were mainly contributed to biological pathways, endocytosis, apoptotic process, trans-Golgi membrane and lysosome, besides, the mTOR, NF-κB , Rap 1, cAMP, TNF, Ras, pathways of cancer and MAPK signaling pathways. What is more, real-time quantitative reverse transcription-polymerase chain reaction verified the expression pictures of ten selected DE miRNAs, which were consistent with the results of the deep sequencing (from 0 hpi up to 48 hpi), and 20 from their corresponding mRNA targets mainly of regulatory pathways of the cellular machinery and immune importance. Our study is the first novel approach which uncover the impact of BRV infection on miRNA expression of MA-104 cells, and it offers clues for identifying potential candidates for antiviral or vaccine strategies.
