Characterization Of MiRNA And Their Target Genes During The Development Of Chicken Skeletal Muscle

MicroRNAs(miRNAs)are non-coding RNAs which can regulate the expression of mRNAs by degrading mRNA molecules or by inhibiting their translation.To investigate the molecular mechanisms of underlying skeletal muscle development based on differentially expressed genes and miRNAs,interactions were constructed according to target prediction result,and integration analysis of up-regulated genes with down regulated miRNAs while down-regulated genes with up-regulated miRNAs with |log2fold change| ≥ 1.75,P < 0.005.From this study has been identified 475 miRNAs(201 up regulated 274 down regulated),492(199 up regulated,293 down-regulated),493(192 up-regulated 302 down-regulated)differential expressed miRNAs found in E11-Vs-E16,E11-Vs-P1 and E16-Vs-P1 comparison respectively.As well 1968(1048 up regulated,920 down-regulated),3249(1647 up-regulated,1602 down-regulated),1525(774 up-regulated,751 down-regulated)were differentially expressed mRNA identified in E11-Vs-E16,E11-Vs-P1 and E16-Vs-P1 comparison respectively.The mi RNA-mRNA integration analysis showed high number of mRNAs regulated by a small number of miRNAs.Significantly expressed miRNAs 51,77,and 29 were analyzed with |log2fold change| > 1 and(P < 0.05)in E11-Vs-E16,E11-Vs-P1,and E16-Vs-P1 comparison respectively.In other hand three significant mi RNAs(gga-miR-205 a,gga-mi R-1a-3p and gga-miR-499-3p)were identified,that equally shared from three comparison group.The integrative validation analysis,from functional in vitro assays showed that,over expression miR-222 a and miR-126-5p in the DF-1 cells,a significant reduction CPEB3(P < 0.01)and FGFR3(P < 0.05)target gene expression level respectively.MiRNA and mRNA RT-qPCR validation result was indicated similar patterns of expression level with RNA deep sequence.Furthermore,gene-ontology functional enrichment analysis of the negative correlation of miRNA-target gene pairs made it important to indicate putative functions of miRNAs in skeletal muscle development.Such as in biological processes,muscle maintenance,myoblast proliferation,muscle thin filament in E11-Vs-E16,comparison group.E11-Vs-P1 comparison,the significant differential expressed genes targeted by miRNA in muscle development,including negative regulation of axon extension,sarcomere organization,and cell redox homeostasis and kinase inhibitor activity identified in biological process.While in E16-Vs-P1 comparison group,negatively regulation of anti-apoptosis,glomerular basement membrane development,negative regulation of axon extension,were identified in biological process.KEGG pathway result suggested that numerous biological pathways were significantly enriched on the oxidative phosphorylation,pyruvate metabolism,phenylalanine,tyrosine and tryptophan biosynthesis,wnt signaling pathway identified from three comparisons group.