Abstract
Noncoding RNAs, especially microRNAs (miRNAs), have been reported to play important roles during skeletal muscle development and regeneration. Our previous sequencing data revealed that miR-99a-5p is one of the most abundant miRNAs in chicken breast muscle. The purpose of this study was to reveal the regulatory mechanism of miR-99a-5p in the proliferation and differentiation of chicken skeletal muscle satellite cells (SMSCs). Through the investigation of cell proliferation activity, cell cycle progression, and 5-ethynyl-29-deoxyuridine (EdU) assay, we found that miR-99a-5p can significantly promote the proliferation of SMSCs. Moreover, we found that miR-99a-5p can inhibit myotube formation by decreasing the expression of muscle cell differentiation marker genes. After miR-99a-5p target gene scanning, we confirmed that miR-99a-5p directly targets the 3′ untranslated region (UTR) of myotubularin-related protein 3 (MTMR3) and regulates its expression level during chicken SMSC proliferation and differentiation. We also explored the role of MTMR3 in muscle development and found that its knockdown significantly facilitates the proliferation but represses the differentiation of SMSCs, which is opposite to the effects of miR-99a-5p. Overall, we demonstrated that miR-99a-5p regulates the proliferation and differentiation of SMSCs by targeting MTMR3.
Highlights
Skeletal muscle is one of the most dynamic and abundant tissues in animals, playing a vital role in movement and metabolism, and accounting for approximately 40% of adult body weight
MiR-99a-5p was enriched in chicken brain (Figure 1C), while being abundant in chicken breast muscle compared with other miRNAs, including the muscle-related miRNAs miR-30a-3p [16] and miR-199-3p [17] (Figure 1D)
To investigate the role of miR-99a-5p in skeletal muscle satellite cells (SMSCs) proliferation and differentiation, cells were transfected with inhibitors or mimics at the proliferation stage or differentiation stage
Summary
Skeletal muscle is one of the most dynamic and abundant tissues in animals, playing a vital role in movement and metabolism, and accounting for approximately 40% of adult body weight. The amount of skeletal muscle determines the meat yield of farm animals. The process of skeletal muscle satellite cell (SMSC) proliferation and differentiation is regulated by many signaling pathways, genes, and noncoding RNAs (ncRNAs) [3,4,5]. An increasing number of studies have shown that miRNAs are involved in skeletal muscle growth and regeneration. MiR-1, Genes 2020, 11, 369; doi:10.3390/genes11040369 www.mdpi.com/journal/genes miR-206, and miR-133 have been identified as being able to regulate skeletal muscle development [8,9]
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