MicroRNA-381 Regulates Proliferation and Differentiation of Caprine Skeletal Muscle Satellite Cells by Targeting PTEN and JAG2

Abstract

In our previous study, microRNA (miR)-381 was found to be the most down-regulated miRNA in skeletal muscle of Liaoning cashmere goats with higher skeletal muscle mass, but the molecular mechanism involved remains unclear. In this study, primary caprine skeletal muscle satellite cells (SMSCs) were isolated and identified. We investigated the effect of miR-381 on the viability, proliferation and differentiation of caprine SMSCs, and the target relationships of miR-381 with jagged canonical Notch ligand 2 (JAG2) and phosphatase and tensin homolog (PTEN). Cells isolated were positive for SMSC-specific marker protein Pax7. This suggests that purified SMSCs were obtained. The expression level of miR-381 achieved a peak value on day 4 after SMSC differentiation, and miR-381 also significantly increased the expression levels of myogenic differentiation marker genes: myosin heavy chain (MyHC), myogenin (MyoG) and myocyte enhancer factor 2C (MEF2C) in differentiated SMSCs, the area of MyHC-positive myotubes and the myogenic index. These findings suggest that miR-381 promoted myogenic differentiation of caprine SMSCs. The CCK8 assay and EDU staining analysis showed that miR-381 mimic both inhibited the viability of SMSCs and decreased the percentage of EDU-labeled positive SMSCs. In contrast, miR-381 inhibitor had the opposite effect with miR-381 mimic. A dual luciferase reporter assay verified that miR-381 can target JAG2 and PTEN by binding to the 3′-untranslated regions (3′-UTR) of the genes. The transfection of miR-381 mimic into caprine SMSCs resulted in decreases in expression levels of JAG2 and PTEN, while miR-381 inhibitor increased the two target genes in expression. This is the first study to reveal the biological mechanisms by which miR-381 regulates caprine SMSC activities.

Transcriptome analysis of mRNA and miRNA in the development of LeiZhou goat muscles

“…We conducted Gene Ontology (GO) enrichment analysis on the target gene candidates of differentially expressed miRNAs. For Gene Ontology (GO) enrichment analysis, we employed a Wallenius non-central hypergeometric distribution based on GOseq 23 , which can adjust for gene length bias. KEGG 24 is a database resource for inferring high-level functions and utilities of biological systems such as the cell, organism, and ecosystem from molecular-level data, especially large-scale molecular datasets generated by genome sequencing and other high-throughput experimental technologies ( http://www.genome.jp/kegg/ ).…”

MicroRNA-542-3p targets Pten to inhibit the myoblasts proliferation but suppresses myogenic differentiation independent of targeted Pten

“…Shen et al . found that Pten positively regulates the proliferation of skeletal muscle satellite cell [ 20 ]. Firstly, the function of Pten regulation on myoblast proliferation was confirmed mediated by si- Pten treatment.…”

Different levels of autophagy induced by transient serum starvation regulate metabolism and differentiation of porcine skeletal muscle satellite cells

“…A certain degree of autophagy helps cells with repair and regeneration, while excessive autophagy has a negative effect on cell function and can lead to cell apoptosis 18 . SMSCs are myogenic stem cells with proliferation and differentiation capabilities after injury or stimulation by certain factors 19 . At present, there is evidence that autophagy can regulate the differentiation of SMSCs 20 , 21 .…”