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Asian-Australas J Anim Sci > Accepted Articles
DOI: https://doi.org/10.5713/ajas.17.0407    [Accepted] Published online January 30, 2018.
Molecular and functional characterization of the adiponectin (AdipoQ) gene in goat skeletal muscle satellite cells
Linjie Wang1, Ke Xue1, Yan Wang1, Lili Niu1, Li Li1, Tao Zhong1, Jiazhong Guo1, Jing Feng2, Tianzeng Song2, Hongping Zhang1,*
1Sichuan Agricultural University, Chengdu, China
2Tibet Academy of Agricultural & Animal Husbandry Science, Lhasa, China
Correspondence:  Hongping Zhang, Tel: 08352882232, Fax: 08352882232, Email: zhp@sicau.edu.cn
Received: 27 May 2017   • Revised: 1 September 2017   • Accepted: 30 January 2018
Abstract
Abstract: Objective: It is commonly accepted that adiponectin binds to its two receptors to regulate fatty acid metabolism in adipocytes. To better understand their functions in the regulation of intramuscular adipogenesis in goats, we cloned the three genes (AdipoQ, AdipoR1 and AdipoR2) encoding these proteins and detected their mRNA distribution in different tissues. We also determined the role of AdipoQ in the adipogenic differentiation of goat skeletal muscle satellite cells (SMSCs).

Methods

SMSCs were isolated using 1 mg/ml Pronase E from the longissimus dorsi muscles of 3-day-old female Nanjiang brown goats. Adipogenic differentiation was induced in satellite cells by transferring the cells to Dulbecco's modified Eagle's medium (DMEM) supplemented with an isobutylmethylxanthine, dexamethasone and insulin (MDI) cocktail. The pEGFP-N1-AD plasmid was transfected into SMSCs using Lipofectamine 2000. Expression of adiponectin in tissues and SMSCs was detected by quantitative polymerase chain reaction (qPCR) and immunocytochemical staining.

Results

The three genes were predominantly expressed in adipose and skeletal muscle tissues. According to fluorescence and immunocytochemical analyses, adiponectin protein expression was only observed in the cytoplasm, suggesting that adiponectin is localized to the cytoplasm of goat SMSCs. In SMSCs overexpressing the AdipoQ gene, adiponectin promoted MSC differentiation into adipocytes and significantly (P < 0.05) up-regulated expression of AdipoR2, ACC, FASN and SREBP-1, though expression of C/EBPα, PPARγ and AdipoR1 did not change significantly.
Conclusion
Adiponectin induced MSC differentiation into adipocytes, indicating that adiponectin may promote intramuscular adipogenesis in goat MSCs.
Keywords: Adiponectin; Intramuscular adipogenesis; Skeletal muscle satellite cells; Fatty acid synthesis


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