Abstract
The fusion of myoblasts is an important step during skeletal muscle differentiation. A recent study in mice found that a transmembrane protein called Myomaker, which is specifically expressed in muscle, is critical for myoblast fusion. However, the cellular mechanism of its roles and the regulatory mechanism of its expression remain unclear. Chicken not only plays an important role in meat production but is also an ideal model organism for muscle development research. Here, we report that Myomaker is also essential for chicken myoblast fusion. Forced expression of Myomaker in chicken primary myoblasts promotes myoblast fusion, whereas knockdown of Myomaker by siRNA inhibits myoblast fusion. MYOD and MYOG, which belong to the family of myogenic regulatory factors, can bind to a conserved E-box located proximal to the Myomaker transcription start site and induce Myomaker transcription. Additionally, miR-140-3p can inhibit Myomaker expression and myoblast fusion, at least in part, by binding to the 3ʹ UTR of Myomaker in vitro. These findings confirm the essential roles of Myomaker in avian myoblast fusion and show that MYOD, MYOG and miR-140-3p can regulate Myomaker expression.
Highlights
Skeletal muscle, which composes approximately half of total body mass, is an important tissue involved in regulating the metabolism, locomotion and strength of the animal body [1]
The results showed that the overexpression of either MYOG or MYOD could significantly increase 0.6 kb promoter activity but that the E-box 1 mutation could eliminate this increase (Figure 5H), suggesting that MYOG and MYOD promote gene transcription by directly binding to the E-box 1 of the Myomaker
The results showed that the overexpression of either MYOG or MYOD could significantly increase 0.6 kb promoter activity but that the E-box 1 mutation could eliminate this increase (Figure 5H), suggesting that MYOG and MYOD promote gene transcription by directly binding to the E-box 1 of the Myomaker promoter
Summary
Skeletal muscle, which composes approximately half of total body mass, is an important tissue involved in regulating the metabolism, locomotion and strength of the animal body [1]. Recent advances in this field have revealed many molecules and signalling pathways that are involved in this process [3] Among these regulatory molecules, transmembrane proteins, which are a type of membrane protein that spans the entirety of the biological membrane, play important roles during myoblast fusion. MYOG and MYOD are critical transcription factors in myogenesis and can regulate the transcription of most of the muscle-specific genes [14,15,16,17] Both of them play an important role in the regulation of myoblast differentiation. We found the regulatory role of MYOG and MYOD in the transcription of Myomaker, and report the expression pattern of these genes during chicken embryonic skeletal muscle development and the differentiation of primary myoblast. Our results confirmed the important roles of Myomaker in avian myoblast fusion and found that MYOD, MYOG and miR-140-3p could regulate Myomaker expression
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