Dystrophin-glycoprotein complex regulates muscle nitric oxide production through mechanoregulation of AMPK signaling.
Patients deficient in dystrophin, a protein that links the cytoskeleton to the extracellular matrix via the dystrophin-glycoprotein complex (DGC), exhibit muscular dystrophy, cardiomyopathy, and impaired muscle nitric oxide (NO) production. We used live-cell NO imaging and in vitro cyclic stretch of isolated adult mouse cardiomyocytes as a model system to investigate if and how the DGC directly regulates the mechanical activation of muscle NO signaling. Acute activation of NO synthesis by mechanical stretch was impaired in dystrophin-deficient mdx cardiomyocytes, accompanied by loss of stretch-induced neuronal NO synthase (nNOS) S1412 phosphorylation. Intriguingly, stretch induced the acute activation of AMP-activated protein kinase (AMPK) in normal cardiomyocytes but not in mdx cardiomyocytes, and specific inhibition of AMPK was sufficient to attenuate mechanoactivation of NO production. Therefore, we tested whether direct pharmacologic activation of AMPK could bypass defective mechanical signaling to restore nNOS activity in dystrophin-deficient cardiomyocytes. Indeed, activation of AMPK with 5-aminoimidazole-4-carboxamide riboside or salicylate increased nNOS S1412 phosphorylation and was sufficient to enhance NO production in mdx cardiomyocytes. We conclude that the DGC promotes the mechanical activation of cardiac nNOS by acting as a mechanosensor to regulate AMPK activity, and that pharmacologic AMPK activation may be a suitable therapeutic strategy for restoring nNOS activity in dystrophin-deficient hearts and muscle.
Department of Molecular and Integrative Physiology, University of Michigan, Ann Arbor, MI 48109;
Joanne F
JF
Garbincius
Department of Molecular and Integrative Physiology, University of Michigan, Ann Arbor, MI 48109; Department of Internal Medicine, University of Michigan, Ann Arbor, MI 48109 dmichele@umich.edu.
Daniel E
DE
Michele
D000263:Adenylate Kinase / Q000378:metabolism*; D000818:Animals; D016189:Dystrophin / Q000378:metabolism*; D004789:Enzyme Activation; D006023:Glycoproteins / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D032383:Myocytes, Cardiac / Q000201:enzymology / Q000378:metabolism*; D009569:Nitric Oxide / Q000096:biosynthesis*; D052248:Nitric Oxide Synthase Type I / Q000378:metabolism; D010766:Phosphorylation; D015398:Signal Transduction
D016189:Dystrophin; D006023:Glycoproteins; D009569:Nitric Oxide; D052248:Nitric Oxide Synthase Type I; D000263:Adenylate Kinase
AMPK; cardiomyocyte; dystrophin; nNOS; stretch