Project description:To characterize the changes of miRNAs in skeletal muscle responding to heat stress, the miRNA expression profiles of longissimus dorsi muscles of pigs raised under constant heat stress (30°C, n=8) or control temperature (22 C, n=8) for 21 days were analyzed by Illumina deep sequencing. 58 differentially expressed miRNAs were identified with 30 down-regulated and 28 up-regulated and 63 differentially expressed target genes were predicted by miRNA-mRNA joint analysis. GO and KEGG analyses showed that the genes regulated by differentially expressed miRNAs were enriched in glucose metabolism, cytoskeletal structure and function, and stress response. Real-time PCR showed that the mRNA levels of PDK4, HSP90 and DES were significantly increased whereas those of SCD and LDH-A significantly decreased by heat exposure. The protein levels of CALM, DES and HIF1α were also significantly increased by constant heat. These results demonstrated that the change of miRNA expression in porcine longissimus dorsi muscle underlie the changes of muscle structure and metabolism in porcine skeletal muscle affected by constant heat stress. Characterize and compare expression profiling of the miRNA transcriptome in 2 groups of porcine skeletal muscle.
Project description:To identify miRNAs involved in muscle development, three independent small RNA libraries from porcine skeletal muscle at the age of 63d、98d and 161d were constructed. A total of 4105184, 4606375 and 5113984 mappable small RNA sequences were generated in the three small RNA libraries, respectively, and 887 known and 472 new candidate miRNAs were obtained.
Project description:The skeletal muscle growth and development is a very complicated but precisely regulated process with interwoven molecular mechanisms. Skeletal muscle is a very heterogeneous tissue that is made up of a large variety of functionally diverse fiber types. Muscle mass is therefore largely determined by the number and size of those fibres. These fibre characteristics are determined by hyperplasia before birth and by hypertrophy after. Around 65 dpc and three postnatal stages (newborn, 3 days; young, 60 days; and mature, 120 days) are key time points in swine skeletal muscle growth and development. We used microarrays to detail the global programme of gene expression underlying porcine skeletal muscle growth and development. Porcine longissimus dorsi muscles were selected at four stages of development for RNA extraction and hybridization on Affymetrix microarrays. We sought to investigate the global gene expression patterns accompanying the skeletal muscle development. To that end, we selected longissimus dorsi muscles at four time-points: 65 days post coitus, 3 days, 60 days and 120 days afterbirth.
Project description:Porcine satellite cells play a vital role in the construction, development, and self-renewal of skeletal muscle. In this study, porcine satellite cells were exposed to mimic viral infection poly (I:C) during proliferation and differentiation phases at 0h, 12h, 24h and 48h time points. The untreated and treated porcine satellite cells during proliferation and differentiation phases were further analyzed by RNA sequencing technology. In the proliferation and differentiation phases of porcine satellite cells grown under poly (I:C), 88, 119, 104 and 95 genes were differentially expressed in 0h – 12h treated, 12h – 24h treated, 0h – 24h treated and 24h – 48h untreated comparison libraries, respectively. The GO terms analysis results showed that in the proliferation phase of treated porcine satellite cells, the up-regulated genes related to the immune system were highly expressed. In addition, the gene expression associated with muscle structure development, response to growth factor emerged in the differentiation phase of untreated porcine satellite cells. The biological pathways associated with Influenza A, Toll-like signaling as well as chemokine signaling were revealed through poly (I:C) stimulation of porcine satellite cells. The differentially expressed genes were confirmed by quantitative real-time PCR. Our findings expanded the understanding of gene expression and signaling pathways about the infiltrated mechanism of the virus into porcine skeletal muscle satellite cells.
Project description:To characterize the mechanism of porcine skeletal muscle development, transcriptome analysis of longissimus dorsi muscle between Shaziling and Yorkshire pig breeds
Project description:We isolated and cultured porcine skeletal muscle satellite cells (PSCs). PSCs were induced to differentiate. At 24h and 36h of differentiation, the PSCs were collected for RNA-seq. Each group had three biological replicates.