A role for calcium-calmodulin in regulating nitric oxide production during skeletal muscle satellite cell activation

• Published in: American Journal of Physiology: Cell Physiology 2009-04, Vol.296 (4), p.C922-C929
• Main Authors: Tatsumi, Ryuichi, Wuollet, Adam L, Tabata, Kuniko, Nishimura, Shotaro, Tabata, Shoji, Mizunoya, Wataru, Ikeuchi, Yoshihide, Allen, Ronald E
• Format: Article
• Language: English
• Subjects: Animals; Binding sites; Calcimycin - pharmacology; Calcium; Calcium - metabolism; Calmodulin - antagonists & inhibitors; Calmodulin - metabolism; Cell culture; Cell cycle; Cell Proliferation - drug effects; Cells, Cultured; Culture Media, Conditioned - metabolism; Dose-Response Relationship, Drug; Enzyme Inhibitors - pharmacology; Hepatocyte Growth Factor - metabolism; Imidazoles - pharmacology; Ionophores - pharmacology; Male; Musculoskeletal system; NG-Nitroarginine Methyl Ester - pharmacology; Nitric oxide; Nitric Oxide - metabolism; Nitric Oxide Synthase - antagonists & inhibitors; Nitric Oxide Synthase - metabolism; Proteins; Rats; Rats, Sprague-Dawley; Satellite Cells, Skeletal Muscle - drug effects; Satellite Cells, Skeletal Muscle - metabolism; Signal Transduction - drug effects; Sulfonamides - pharmacology
• ISSN: 0363-6143; 1522-1563
• DOI: 10.1152/ajpcell.00471.2008

1 Department of Bioscience and Biotechnology and 3 Department of Animal Science, Graduate School of Agriculture, Kyushu University, Fukuoka, Japan; 2 Muscle Biology Group, Department of Animal Sciences, College of Agriculture and Life Sciences, University of Arizona, Tucson, Arizona Submitted 12 September 2008 ; accepted in final form 19 January 2009 When skeletal muscle is stretched or injured, myogenic satellite cells are activated to enter the cell cycle. This process depends on nitric oxide (NO) production by NO synthase (NOS), matrix metalloproteinase activation, release of hepatocyte growth factor (HGF) from the extracellular matrix, and presentation of HGF to the c-met receptor as demonstrated by a primary culture and in vivo assays. We now add evidence that calcium-calmodulin is involved in the satellite cell activation cascade in vitro. Conditioned medium from cultures that were treated with a calcium ionophore (A23187 [GenBank] , ionomycin) for 2 h activated cultured satellite cells and contained active HGF, similar to the effect of mechanical stretch or NO donor treatments. The response was abolished by addition of calmodulin inhibitors (calmidazolium, W-13, W-12) or a NOS inhibitor N G -nitro- L -arginine methyl ester hydrochloride but not by its less inactive enantiomer N G -nitro- D -arginine methyl ester hydrochloride. Satellite cells were also shown to express functional calmodulin protein having a calcium-binding activity at 12 h postplating, which is the time at which the calcium ionophore was added in this study and the stretch treatment was applied in our previous experiments. Therefore, results from these experiments provide an additional insight that calcium-calmodulin mediates HGF release from the matrix and that this step in the activation pathway is upstream from NO synthesis. muscle regeneration; stretch-activation Address for reprint requests and other correspondence: R. E. Allen, Muscle Biology Group, Dept. of Animal Sciences, Coll. of Agriculture and Life Sciences, Univ. of Arizona, Shantz Blvd. 204, PO Box 210038, Tucson, AZ 85721-0038 (e-mail: rallen{at}ag.arizona.edu )