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Effect of MSTN Knockdown by Antisense RNA on Expression of Genes Associated with Fat Deposition in Bovine (Bos taurus) Muscle Satellite Cells |
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Abstract Myostatin (MSTN) is a member of the transforming growth factor β (TGF-β) superfamily, which plays a key role in muscle development. Numerous studies have shown that knockout of MSTN results in a widespread increase in skeletal muscle mass, and a significant reduction in fat accumulation. However, the mechanisms involved in fat deposition process yielded by MSTN gene suppression has not been understood. In the present study, we constructed 5 two-way expression antisense RNA vectors pMSTN-CMV-CAG-antiMSTN, then transfected bovine fetal fibroblast cells, and screened the best antisense RNA vector which significantly disturbed the MSTN expression in protein level with Western blot. The vectors were then confected to a bovine muscle satellite cells and the effect of MSTN knockdown to genes associated with adipogenesis such as fatty acid synthase (FASN), acetyl-CoA carboxylase (ACC), stearoyl-CoA desaturase-1 (SCD-1), genes associated with lipolysis such as carnitine palmitoyltransferase 1b (CPT-1B), acyl-CoA oxidase(ACOX1) and enoyl-CoA hydratase 1 (ECHDC1), and fat transport protein (FATP) gene were investigated. The results showed that genes associated with adipogenesis and lipolysis were all up-regulated except the expression of ECHDC1 when MSTN knockdown in muscle satellite cells. The expression of gene associated with fat transport has no significant change. The results suggested that MSTN knockdown could enhance both adipogenesis and lipolysis, so the reduced fat mass resulted from MSTN knockdown was due to the increased lipolysis rather than inhibition of adipogenesis. And the expression of CPT-1B was up-regulated in the highest level among the genes investigated, this result suggested that the effect of MSTN to fat was mainly by the process of β oxidation, because CPT-1B was a main factor of β oxidation. This study provides a theoretical basis for the decreased fat content resulted from MSTN reduction, and makes a preliminary study for the effect of MSTN on fat deposition.
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Received: 20 June 2016
Published: 07 November 2016
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