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MiRNA-seq of lung in two kinds of pig breeds after PCV2 injection

ABSTRACT: Postweaning multisystemic wasting syndrome in pigs has devastated the swine industry since the 1990s. Porcine circovirus type 2 (PCV2) is the primary cause of this disease. MicroRNAs (miRNAs) are small noncoding RNAs that play important roles in regulating gene expression, especially at the post-transcription level. The expression profiles of miRNAs have been reported in porcine reproductive and respiratory syndrome, porcine parvovirus, and other pig diseases; however, the miRNA expression profiles in pigs infected with PCV2 have not been reported so far. The Laiwu pig (a Chinese indigenous pig breed) has different meat quality, adipogenesis, and disease-resistance from western commercial pig breeds. In this study, four small RNA libraries were constructed from the lung tissue of uninfected and infected Laiwu and Yorkshire/Landrace crossbred (YL) pigs. High-throughput sequencing and bioinformatics were used to determine the abundance and differential expression of miRNAs in the four libraries

ORGANISM(S): Sus scrofa

PROVIDER: GSE92259 | GEO | 2016/12/13

SECONDARY ACCESSION(S): PRJNA357045

REPOSITORIES: GEO

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Project description:Purpose: Post-weaning multisystemic wasting syndrome (PMWS) is a disease associated with porcine circovirus type 2 (PCV2) infection. One main feature of PMWS is seriously decreased lymphocyte in lymphoid tissues. This research aims to investigate the molecular mechanism of evident lymphocyte depletion in spleen from Yorkshire x Landrace (YL) pigs after infection with PCV2. Methods: At the age of 6 weeks, 15 piglets of Laiwu (LW) and YL pigs were assigned randomly into two groups, respectively: 10 infected pigs and 5 uninfected pigs. Spleen tissues of each group were collected at 35 days post infection with PCV2. Three PCV2-infected YL pigs with apparently pathologic characteristics and mock-infected YL pigs were selected for RNA sequencing, respectively. The RNA extraction, library construction and RNA-Seq were performed on the Illumina HiSeq2500 at Beijing BioMarker Technologies (Beijing, China).Differential expression genes (DEGs) were filtrated from the result of transcriptome. The quantitative real-time PCR was performed to validate the expression patterns of candidate DEGs between LW and YL pigs. Results: The clean reads percolated from the raw reads were mapped to Sus scrofa genome (Sscrofa10.2) using Tophat2 software. The efficiency of alignment was about 76% between the six samples and reference genome, and approximate 50% clean reads were perfectly matched to pig genome. Two samples with high R-squared value (T1-T3: 0.98; T5-T6: 0.97) from PCV2 and mock-infected YL pigs, respectively, was performed differential expression analysis through DESeq after biologic repetition correlation detection. With the criterion of log FC (fold change) > 1.5 and of P value < 0.05, 90 significantly differentially expressed genes were identified between the two groups, of which 57 were up-regulated and 33 were down-regulated. Eleven candidate DEGs were confirmed by quantitative real-time PCR, which expression pattern were generally in accordance with the results of RNA-seq. Conclusions: Many DEGs participate in immune response and some of them are involved in cells proliferation or apoptosis, such as CD81, HGF, CXCL13, KLF11, PSMD4, CYCS and ACTC1. The expression changes of these DEGs may cause the lymphocyte depletion in spleen after challenge with PCV2.

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MiRNA-seq of lung in two kinds of pig breeds after PCV2 injection

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