MiR-194-5p negatively regulates the proliferation and differentiation of rabbit skeletal muscle satellite cells

Yu Shi,Songjia Lai,Xiulan Lai,Shenqiang Hu,Shiyi Chen,Xianbo Jia,Jie Wang,Mingcheng Cai,Xudong Mao

doi:10.1007/s11010-020-03918-0

Abstract

Skeletal muscle satellite cells (SMSCs), also known as a multipotential stem cell population, play a crucial role during muscle growth and regeneration. In recent years, numerous miRNAs have been associated with the proliferation and differentiation of SMSCs in a number of mammalian species; however, the regulatory mechanisms of miR-194-5p in rabbit SMSCs still remain scarce. In this study, miR-194-5p was first observed to be highly expressed in the rabbit leg muscle. Furthermore, both the mimics and inhibitor of miR-194-5p were used to explore its role in the proliferation and differentiation of rabbit SMSCs cultured in vitro. Results from both EdU and CCK8 assays showed that miR-194-5p inhibited the proliferation of SMSCs. Meanwhile, Mef2c was identified as a target gene of miR-194-5p based on the dual-luciferase reporter assay results. In addition, upregulation of miR-194-5p decreased the expression levels of Mef2c and MyoG during rabbit SMSCs differentiation on Days 3 and 7 of in vitro culture. Taken together, these data demonstrated that miR-194-5p negatively regulates the proliferation and differentiation of rabbit SMSCs by targeting Mef2c.

Highlights

Skeletal muscle satellite cells (SMSCs), known as a multipotential stem cell population, mainly exist on the surface of myofiber but beneath the basement membrane
We aimed to investigate the expression of miR-194-5p in the leg muscle of rabbit and reveal the possible role of miR-194-5p in the proliferation and differentiation of rabbit SMSCs through gain and loss function of miR-194-5p
To further verify the relationship between Mef2c and miR-194-5p, the expression of Mef2c in rabbit SMSCs was measured after transfection of miR-194-5p mimics, NC, inhibitor, and inNC, and the results showed that over-expression of miR-194-5p significantly suppressed the Mef2c expression, but there was no significant difference between the treatments of miR-194-5p inhibitor and inNC (Fig. 5)

Summary

Introduction

Skeletal muscle satellite cells (SMSCs), known as a multipotential stem cell population, mainly exist on the surface of myofiber but beneath the basement membrane. Adult SMSCs are quiescent, but they are prone to be activated by physical trauma or growth signals [1] Over the last few decades, many studies have proved that miRNAs play important roles in a variety of cellular activities, including cell proliferation, differentiation, and apoptosis [7, 8]. Lots of studies have indicated that miRNAs play a role in myogenesis.

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