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MiR-188-5p regulates the proliferation and differentiation of goat skeletal muscle satellite cells by targeting calcium/calmodulin dependent protein kinase II beta |
Jing Jing1,2,a 
, Sihuan Zhang1,2,a
, Jinbo Wei1,2
, Yuhang Yang1,2
, Qi Zheng1,2
, Cuiyun Zhu1,2
, Shuang Li1,2
, Hongguo Cao1,2
, Fugui Fang1,2
, Yong Liu3
, Ying-hui Ling1,2,3,*
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1College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China
2Anhui Province Key Laboratory of Local Livestock and Poultry Genetic Resource Conservation and Bio-Breeding, Anhui Agricultural University, Hefei 230036, China
3Key Laboratory of Embryo Development and Reproductive Regulation of Anhui Province, Fuyang Normal University, Fuyang, Anhui Province 236041, China |
Correspondence:
Ying-hui Ling, Tel: +86-551-65785928, Fax: +86-551-65785928, Email: lingyinghui@ahau.edu.cn
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Received: 13 March 2023 • Revised: 29 April 2023 • Accepted: 14 June 2023
aThese authors contributed equally to this work. |
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| Abstract |
Objective
The aim of this study was to reveal the role and regulatory mechanism of miR-188-5p in the proliferation and differentiation of goat muscle satellite cells.
Methods
Goat skeletal muscle satellite cells isolated in the pre-laboratory were used as the test material. First, the expression of miR-188-5p in goat muscle tissues at different developmental stages was detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR). In addition, miR-188-5p was transfected into goat skeletal muscle satellite cells by constructing mimics and inhibitors of miR-188-5p, respectively. The changes of differentiation marker gene expression were detected by qPCR method.
Results
It was highly expressed in adult goat latissimus dorsi and leg muscles, goat fetal skeletal muscle, and at the differentiation stage of muscle satellite cells. Overexpression and interference of miR-188-5p showed that miR-188-5p inhibited the proliferation and promoted the differentiation of goat muscle satellite cells. Target gene prediction and dual luciferase assays showed that miR-188-5p could target the 3'untranslated region of the calcium/calmodulin dependent protein kinase II beta (CAMK2B) gene and inhibit luciferase activity. Further functional studies revealed that CAMK2B promoted the proliferation and inhibited the differentiation of goat muscle satellite cells, whereas si-CAMK2B restored the function of miR-188-5p inhibitor.
Conclusion
These results suggest that miR-188-5p inhibits the proliferation and promotes the differentiation of goat muscle satellite cells by targeting CAMK2B. This study will provide a theoretical reference for future studies on the molecular mechanisms of skeletal muscle development in goats. |
| Keywords:
CAMK2B; Goat; miR-188-5p; Myogenic Differentiation; Myogenic Proliferation; Skeletal Muscle Satellite Cells |
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