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| Effects of miR-133a on the Proliferation and Differentiation of Bovine (Bos taurus) Skeletal Muscle-derived Satellite Cells |
| LI Yan1, YANG Xu1, CHEN Ming-Ming1, GUO Hong1, DING Xiang-Bin1, LIU Xin-Feng1,2,3,* |
1 Tianjin Key Laboratory of Agricultural Animal Breeding and Healthy Husbandry/College of Animal Science and Veterinary Medicine, Tianjin Agricultural University, Tianjin 300384, China;
2 Key Laboratory of Education for Conservation and Utilization of Special Biological Resources in the Western China, Ningxia University, Yinchuan 750021, China;
3 School of Life Science, Ningxia University, Yinchuan 750021, China |
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Abstract miR-133a is the microRNA (miRNA) specifically expressed in skeletal muscle and myocardium, but its role in the proliferation and differentiation of skeletal muscle-derived satellite cells (MDSCs) in Bos taurus is unknown. In order to further understand the effect of miR-133a on the proliferation and differentiation in bovine MDSCs, and enrich the molecular genetic mechanism affecting the growth and development of bovine muscle, in this study, based on the established in vitro differentiation model of Bos taurus MDSCs, qRT-PCR method was used to detect the temporal expression profiles of miR-133a, myogenin (MyoG), myogenic differentiation (MyoD) and myosin heavy chain (MHC) in MDSCs. Then mimic and inhibitors of miR-133a were used to transfect BMSCs to construct a model of miR-133a overexpressing or inhibiting, and transfection effects on MDSCs proliferation were detected by 5-ethynyl-2'-deoxyuridine (EdU) staining for proliferating cells. The techniques of qRT-PCR, Western blot and immunofluorescence were used to detect the mRNA and protein levels of MyoG, MyoD and MHC. The results showed that the expression level of miR-133a increased significantly at the differentiation stage of MDSCs (P<0.01). After overexpression of miR-133a, the positive ratio of proliferating cells significantly decreased (P<0.01), while the gene expression of MyoG (P<0.05) and MHC (P<0.01) significantly increased; The protein expression level of MHC significantly increased (P<0.01) and the number of fused myotubes increased. After miR-133a expression was inhibited, the positive ratio of proliferating cells significantly increased (P<0.01), while the gene expression of MyoG (P<0.01), MyoD (P<0.05) and MHC (P<0.01) significantly decreased; the expression of MHC protein significantly decreased (P<0.01) and the number of fused myotubes decreased. The above results confirmed that miR-133a played a role in inhibiting proliferation and promoting differentiation of MDSCs. The present study enriches the molecular genetic research of bovine muscle growth and development, and provides an important reference for further research on the function of miRNA in regulating animal muscle growth and development.
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Received: 22 August 2020
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Corresponding Authors:
* liu2019074@nxu.edu.cn
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