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 characterize the mechanism of porcine skeletal muscle development, transcriptome analysis of longissimus dorsi muscle between Shaziling and Yorkshire pig breeds
Project description:Solexa/Illumina deep sequencing was used to obtain miRNA transcriptome profiles for control (basal diet, 14.49% crude protein, 13.73 MJ/kg digestible energy, 0.86% available lysine) and traditional Chinese medicine formula (basal diet+2.5 g/kg traditional Chinese medicine formula) groups. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analysis was performed using DAVID functional annotation to identify the biological process function of the miRNAs identified as being involved in muscle fiber regulation. We detected 269 mature miRNAs and 14 pre-miRNAs in the porcine muscle samples, of which 211 were previously unreported. Pathway analyses suggested that several independent signaling pathways were involved in the biological process, cellular component, and molecular function categories, with metabolic pathways showing the most enrichment and including some pathways involved in the regulation of muscle fiber phenotype such as mitogen-activated protein kinase (MAPK), mammalian target of rapamycin, and citrate cycle signaling pathways. Thirty-seven miRNAs exhibited normalized expression of over 10,000 reads per million, accounting for 85.91% of the overall counts of the total porcine mature miRNAs. Among these, 9 were muscle-related miRNAs involved in muscle development, including skeletal satellite cell (ssc)-miR-133a-3p, ssc-miR-486, ssc-miR-26a, ssc-miR-21, ssc-miR-1, ssc-miR-10b, ssc-miR-181a, ssc-miR-128, and ssc-miR-23a. In particular, miR-1 and miR-133 are muscle-specific miRNAs, termed myomiRs, which play an important role in muscle differentiation and development. Furthermore, porcine skeletal satellite cell transfection experiments indicated that overexpression or inhibition of ssc-miR-27a repressed or increased, respectively, MYH7 gene and protein expression through the PGC-1-MEF2C pathway, suggesting that miR-27a participated in the regulation of muscle fiber in skeletal muscle. In conclusion, these miRNAome profiles provide novel insight into the mechanism underlying muscle fiber regulation.
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 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.