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MicroRNA bta-miR-365-3p inhibits proliferation but promotes differentiation of primary bovine myoblasts by targeting the activin A receptor type I

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  • Dan Hao
  • Xiaogang Wang, Northwest Agriculture and Forestry University
  • ,
  • Xiao Wang, Danmarks Tekniske Universitet
  • ,
  • Bo Thomsen
  • Yu Yang, Northwest Agriculture and Forestry University
  • ,
  • Xianyong Lan, Northwest Agriculture and Forestry University
  • ,
  • Yongzhen Huang, Northwest Agriculture and Forestry University
  • ,
  • Hong Chen, Northwest Agriculture and Forestry University

Background: MicroRNAs act as post-transcriptional regulators that repress translation or degrade mRNA transcripts. Each microRNA has many mRNA targets and each mRNA may be targeted by several microRNAs. Skeletal muscles express a plethora of microRNA genes that regulate muscle development and function by controlling the expression of protein-coding target genes. To expand our understanding of the role of microRNA, specifically bta-miR-365-3p, in muscle biology, we investigated its functions in regulating primary bovine myoblast proliferation and differentiation. Results: Firstly, we found that bta-miR-365-3p was predominantly expressed in skeletal muscle and heart tissue in Chinese Qinchuan beef cattle. Quantitative PCR and western blotting results showed that overexpression of bta-miR-365-3p significantly reduced the expression levels of cyclin D1 (CCND1), cyclin dependent kinase 2 (CDK2) and proliferating cell nuclear antigen (PCNA) but stimulated the expression levels of muscle differentiation markers, i.e., MYOD1, MYOG at both mRNA and protein level. Moreover, downregulation of bta-miR-365-3p increased the expression of CCND1, CDK2 and PCNA but decreased the expression of MYOD1 and MYOG at both mRNA and protein levels. Furthermore, flow cytometry, EdU proliferation assays and immunostaining results showed that increased levels of bta-miR-365-3p suppressed cell proliferation but promoted myotube formation, whereas decreased levels of bta-miR-365-3p resulted in the opposite consequences. Finally, we identified that activin A receptor type I (ACVR1) could be a direct target of bta-miR-365-3p. It was demonstrated that bta-miR-365-3p can bind to the 3’UTR of ACVR1 gene to regulate its expression based on dual luciferase gene reporter assays. Consistently, knock-down of ACVR1 was associated with decreased expressions of CDK2, CCND1 and PCNA but increased expression of MYOG and MYOD1 both at mRNA and protein level. Conclusion: Collectively, these data suggested that bta-miR-365-3p represses proliferation but promotes differentiation of bovine myoblasts through several biological mechanisms involving downregulation of ACVR1.

OriginalsprogEngelsk
Artikelnummer16
TidsskriftJournal of Animal Science and Biotechnology
Vol/bind12
Nummer1
Antal sider15
ISSN1674-9782
DOI
StatusUdgivet - jan. 2021

Bibliografisk note

Funding Information:
This work was supported by the National Natural Science Foundation of China (No.31772574) the Program of National Beef Cattle and Yak Industrial Technology System (CARS-37). Dan Hao and Xiao Wang appreciated the scholarship from the China Scholarship Council (CSC), China.

Publisher Copyright:
© 2021, The Author(s).

Copyright:
Copyright 2021 Elsevier B.V., All rights reserved.

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