1,25(OH)2-vitamin D3 enhances the stimulating effect of leucine and insulin on protein synthesis rate through Akt/PKB and mTOR mediated pathways in murine C2C12 skeletal myotubes
Article first published online: 9 AUG 2013
© 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Molecular Nutrition & Food Research
Volume 57, Issue 12, pages 2137–2146, December 2013
How to Cite
Salles, J., Chanet, A., Giraudet, C., Patrac, V., Pierre, P., Jourdan, M., Luiking, Y. C., Verlaan, S., Migné, C., Boirie, Y. and Walrand, S. (2013), 1,25(OH)2-vitamin D3 enhances the stimulating effect of leucine and insulin on protein synthesis rate through Akt/PKB and mTOR mediated pathways in murine C2C12 skeletal myotubes. Mol. Nutr. Food Res., 57: 2137–2146. doi: 10.1002/mnfr.201300074
- Issue published online: 4 DEC 2013
- Article first published online: 9 AUG 2013
- Manuscript Accepted: 28 MAY 2013
- Manuscript Revised: 22 APR 2013
- Manuscript Received: 13 FEB 2013
- Nutricia Advanced Medical Nutrition, Danone Research, Centre for Specialised Nutrition, Wageningen, the Netherlands
- National Institute for Agricultural Research
- Akt/mTOR pathway;
- C2C12 myotubes;
- 1,25(OH)2-vitamin D3;
- Protein synthesis rate;
- Skeletal muscle
In recent years, there has been a growing body of evidence pointing to an effect of vitamin D on muscle mass and function. Our aim was to investigate the combined effect of 1,25(OH)2-vitamin D3 (1,25(OH)2D3) with anabolic factors insulin and leucine on protein fractional synthesis rate (FSR) and regulation in the mouse C2C12 myotube.
Methods and results
After differentiation, myotubes were cultured in 1,25(OH)2D3 solutions at 0, 1, or 10 nM for 72 h. Cells were treated by l-[1–13C]valine and puromycin in presence or not of leucine and insulin, and protein FSR was determined by measuring tracer enrichments and puromycin incorporation in proteins, respectively. Protein expression and phosphorylation state of insulin receptor (IR), Akt, GSK3, mTOR, p70 S6 kinase, rpS6, and 4EBP1 were measured by Western blot. Transcript levels of IR and 1,25(OH)2D3 receptor (VDR) were determined by qPCR. 1,25(OH)2D3 (10 nM) with leucine and insulin increased protein FSR in C2C12 myotubes (14–16%). IR and VDR mRNA expression was increased with 1,25(OH)2D3 treatment. The Akt/mTOR-dependent pathway was activated by insulin and leucine and further enhanced by 1,25(OH)2D3.
1,25(OH)2D3 sensitizes the Akt/mTOR-dependant pathway to the stimulating effect of leucine and insulin, resulting in a further activation of protein synthesis in murine C2C12 skeletal myotubes.