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Asian-Australas J Anim Sci > Accepted Articles
DOI: https://doi.org/10.5713/ajas.20.0430    [Accepted] Published online October 13, 2020.
Isolation and identification of goose skeletal muscle satellite cells and preliminary study on the function of CTRP3 gene
Han Wang1  , Ke He1  , Xuehua Zeng1  , Xiaolong Zhou1  , Feifei Yan1  , Songbai Yang1  , Ayong Zhao1,* 
College of Animal Science and Technology • College of Veterinary Medicine, Zhejiang A&F University, Zhejiang 311300, China
Correspondence:  Ayong Zhao, Tel: +86-0571-63741575, Fax: +86-0571-63741575, Email: zay503@zafu.edu.cn
Received: 19 June 2020   • Revised: 17 August 2020   • Accepted: 2 September 2020
Abstract
Objective
Skeletal muscle satellite cells (SMSCs) are significant for the growth, regeneration, and maintenance of skeletal muscle after birth. However, currently, few studies have been performed on the isolation, culture and inducing differentiation of goose muscle satellite cells. Previous studies have shown that CTRP3 participated in the process of muscle growth and development, but its role in the goose skeletal muscle development is not yet clear. This study aimed to isolate, culture, and identify the goose SMSCs in vitro. Additionally, to explore the function of CTRP3 in goose SMSCs.
Methods
Goose SMSCs were isolated using 0.25% trypsin from leg muscle of 15-20 day fertilized goose eggs. Cell differentiation was induced by transferring the cells to differentiation medium (DM) with 2% horse serum and 1% penicillin streptomycin. Immunofluorescence staining of Desmin and Pax7 was used to identify goose SMSCs. Quantitative real­time polymerase chain reaction and western blot were applied to explore developmental expression profile of CTRP3 in LM and the regulation of CTRP3 on myosin heavy chains (MyHC), myogenin (MyoG) expression and Notch signaling pathway related genes expression.
Results
The goose SMSCs were successfully isolated and cultured. The expression of Pax7 and Desmin were observed in the isolated cells. The expression of CTRP3 decreased significantly during leg muscle development. Overexpression of CTRP3 could enhance the expression of two myogenic differentiation marker genes, MyHC and MyoG. But knockdown of CTRP3 suppressed their expression. Furthermore, CTRP3 could repress the mRNA level of Notch signaling pathway-related genes, notch receptor 1 (Notch1), notch receptor 2 (Notch2) and hairy/enhancer-of-split related with YRPW motif 1 (Hey1), which previously showed a negative regulation in myoblast differentiation.
Conclusion
These findings provide a useful cell model for the future research on goose muscle development and suggest that CTRP3 may play an essential role in skeletal muscle growth of goose.
Keywords: Goose; Skeletal Muscle Satellite Cells; CTRP3; Differentiation


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