Numerous physiological and pathological processes are controlled by free fatty acids, which act as signaling molecules in mammals. We hypothesized that oleic acid (Ole) might stimulate the formation of satellite-cell-...Numerous physiological and pathological processes are controlled by free fatty acids, which act as signaling molecules in mammals. We hypothesized that oleic acid (Ole) might stimulate the formation of satellite-cell-derived intramuscular adipose tissue. The role of Ole as a ligand of <em>G-protein-coupled receptor 43</em> (<em>GPR4</em>3) was previously identified. Thus, the objective of the current study was to determine the effect of Ole on <em>GPR43</em> and factors related to the adipogenic differentiation of bovine satellite cells (BSC). Treatments of 100 μM and 500 μM Ole tended to induce greater (P < 0.10) mRNA expression of <em>CCAAT/enhancer-binding protein β</em> (<em>C/EBPβ</em>) compared to all other doses. The mRNA abundance of peroxisome proliferator-activated receptor γ (<em>PPARγ</em>) was not altered (<em>P</em> > 0.10) by treatment. The addition of 100 μM and 500 μM of Ole upregulated (<em>P</em> < 0.05) <em>GPR43</em> mRNA expression. Protein level of GPR43 was increased (<em>P</em> < 0.05) by 100 μM of Ole treatments. Addition of Ole to BSC cultures induced transformation of myogenic cells into adipocyte-like cells that formed cytoplasmic lipid droplets. Increased expression of<em> C/EBPβ</em> in response to Ole might suppress myogenic differentiation. After the treatment of cells with Ole, increased expression of GPR43 could lead to phosphorylation of 5’ AMP-activated protein kinase α (AMPKα). Altogether, the results indicated that increased Ole might stimulate adipose tissue accumulation within the skeletal muscle of cattle by promoting adipogenic differentiation and activation of GPR43 in satellite cells.展开更多
To examine the effect of myogenin gene on the differentiation of bovine skeletal muscle satellite cell, we constructed small interfering RNA plasmid vector to obtain myogenin knockdown bovine skeletal muscle cells, th...To examine the effect of myogenin gene on the differentiation of bovine skeletal muscle satellite cell, we constructed small interfering RNA plasmid vector to obtain myogenin knockdown bovine skeletal muscle cells, then used cell transfection, real time RCR and Western Blot to detect the influence of myogenin to cell differentiation. Results showed that the knockdown of myogenin significantly decreased its expression and other muscle-specific genes. Compared to the control, it could differentiate into few myotubes when challenged by low serum in the medium. These findings provided an important theoretical basis for further explore of the genetic mechanism in adult skeletal muscle, the remedy of muscle injuries and the cultivation of high-yield transgenic cattle.展开更多
文摘Numerous physiological and pathological processes are controlled by free fatty acids, which act as signaling molecules in mammals. We hypothesized that oleic acid (Ole) might stimulate the formation of satellite-cell-derived intramuscular adipose tissue. The role of Ole as a ligand of <em>G-protein-coupled receptor 43</em> (<em>GPR4</em>3) was previously identified. Thus, the objective of the current study was to determine the effect of Ole on <em>GPR43</em> and factors related to the adipogenic differentiation of bovine satellite cells (BSC). Treatments of 100 μM and 500 μM Ole tended to induce greater (P < 0.10) mRNA expression of <em>CCAAT/enhancer-binding protein β</em> (<em>C/EBPβ</em>) compared to all other doses. The mRNA abundance of peroxisome proliferator-activated receptor γ (<em>PPARγ</em>) was not altered (<em>P</em> > 0.10) by treatment. The addition of 100 μM and 500 μM of Ole upregulated (<em>P</em> < 0.05) <em>GPR43</em> mRNA expression. Protein level of GPR43 was increased (<em>P</em> < 0.05) by 100 μM of Ole treatments. Addition of Ole to BSC cultures induced transformation of myogenic cells into adipocyte-like cells that formed cytoplasmic lipid droplets. Increased expression of<em> C/EBPβ</em> in response to Ole might suppress myogenic differentiation. After the treatment of cells with Ole, increased expression of GPR43 could lead to phosphorylation of 5’ AMP-activated protein kinase α (AMPKα). Altogether, the results indicated that increased Ole might stimulate adipose tissue accumulation within the skeletal muscle of cattle by promoting adipogenic differentiation and activation of GPR43 in satellite cells.
基金Supported by the Ministry of Agricultural Nuarture of New Varieties Genetically Modified Organisms Significant Special Funding (2008ZX08007-002)
文摘To examine the effect of myogenin gene on the differentiation of bovine skeletal muscle satellite cell, we constructed small interfering RNA plasmid vector to obtain myogenin knockdown bovine skeletal muscle cells, then used cell transfection, real time RCR and Western Blot to detect the influence of myogenin to cell differentiation. Results showed that the knockdown of myogenin significantly decreased its expression and other muscle-specific genes. Compared to the control, it could differentiate into few myotubes when challenged by low serum in the medium. These findings provided an important theoretical basis for further explore of the genetic mechanism in adult skeletal muscle, the remedy of muscle injuries and the cultivation of high-yield transgenic cattle.
