Journal article
Nitric oxide and AMPK cooperatively regulate PGC-1 alpha in skeletal muscle cells
The Journal of physiology, Vol.588(18), pp.3551-3566
09/15/2010
DOI: 10.1113/jphysiol.2010.194035
PMCID: PMC2988518
PMID: 20643772
Abstract
Nitric oxide (NO) induces mitochondrial biogenesis in skeletal muscle cells via upregulation of the peroxisome proliferator-activated receptor-gamma coactivator 1 alpha (PGC-1 alpha). Further, we have shown that nitric oxide interacts with the metabolic sensor enzyme, AMPK. Therefore, we tested the hypothesis that nitric oxide and AMPK act synergistically to upregulate PGC-1 alpha mRNA expression and stimulate mitochondrial biogenesis in culture. L6 myotubes treated with nitric oxide donors, S-nitroso-N-penicillamine (SNAP, 25 mu m) or diethylenetriamine-NONO (DETA-NO, 50 mu m), exhibited elevated AMPK phosphorylation, PGC-1 alpha mRNA and protein, and basal and uncoupled mitochondrial respiration (P < 0.05). Pre-treatment of cultures with the AMPK inhibitor, Compound C, prevented these effects. Knockdown of AMPK alpha 1 in L6 myotubes using siRNA reduced AMPK alpha protein content and prevented upregulation of PGC-1 alpha mRNA by DETA-NO. Meanwhile, siRNA knockdown of AMPK alpha 2 had no effect on total AMPK alpha protein content or PGC-1 alpha mRNA. These results suggest that NO effects on PGC-1 alpha expression are mediated by AMPK alpha 1. Paradoxically, we found that the AMPK-activating compound, AICAR, induced NO release from L6 myotubes, and that AICAR-induced upregulation of PGC-1 alpha mRNA was prevented by inhibition of NOS with NG-nitro-l-arginine methyl ester (l-NAME, 1 mm). Additionally, incubation of isolated mouse extensor digitorum longus (EDL) muscles with 2 mm AICAR for 20 min or electrical stimulation (10 Hz, 13 V) for 10 min induced phosphorylation of AMPK alpha (P < 0.05), which was completely prevented by pre-treatment with the NOS inhibitor, l-NG-monomethyl arginine (l-NMMA, 1 mm). These data identify the AMPK alpha 1 isoform as the mediator of NO-induced effects in skeletal muscle cells. Further, this study supports a proposed model of synergistic interaction between AMPK and NOS that is critical for maintenance of metabolic function in skeletal muscle cells.
Details
- Title: Subtitle
- Nitric oxide and AMPK cooperatively regulate PGC-1 alpha in skeletal muscle cells
- Creators
- Vitor A. Lira - University of FloridaDana L. Brown - University of FloridaAna K. Lira - University of FloridaAndreas N. Kavazis - University of FloridaQuinlyn A. Soltow - University of FloridaElizabeth H. Zeanah - University of FloridaDavid S. Criswell - University of Florida
- Resource Type
- Journal article
- Publication Details
- The Journal of physiology, Vol.588(18), pp.3551-3566
- DOI
- 10.1113/jphysiol.2010.194035
- PMID
- 20643772
- PMCID
- PMC2988518
- NLM abbreviation
- J Physiol
- ISSN
- 0022-3751
- eISSN
- 1469-7793
- Publisher
- Wiley
- Number of pages
- 16
- Grant note
- 0530196N / AHA; American Heart Association
- Language
- English
- Date published
- 09/15/2010
- Academic Unit
- Health, Sport, and Human Physiology
- Record Identifier
- 9984259645602771
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