title: Intracellular calcium handling in ventricular myocytes from mdx mice.
abstract: Duchenne muscular dystrophy (DMD) is a lethal degenerative disease of skeletal muscle, characterized by the absence of the cytoskeletal protein dystrophin. Some DMD patients show a dilated cardiomyopathy leading to heart failure. This study explores the possibility that dystrophin is involved in the regulation of a stretch-activated channel (SAC), which in the absence of dystrophin has increased activity and allows greater Ca(2+) into cardiomyocytes. Because cardiac failure only appears late in the progression of DMD, we examined age-related effects in the mdx mouse, an animal model of DMD. Ca(2+) measurements using a fluorescent Ca(2+)-sensitive dye fluo-4 were performed on single ventricular myocytes from mdx and wild-type mice. Immunoblotting and immunohistochemistry were performed on whole hearts to determine expression levels of key proteins involved in excitation-contraction coupling. Old mdx mice had raised resting intracellular Ca(2+) concentration ([Ca(2+)](i)). Isolated ventricular myocytes from young and old mdx mice displayed abnormal Ca(2+) transients, increased protein expression of the ryanodine receptor, and decreased protein expression of serine-16-phosphorylated phospholamban. Caffeine-induced Ca(2+) transients showed that the Na(+)/Ca(2+) exchanger function was increased in old mdx mice. Two SAC inhibitors streptomycin and GsMTx-4 both reduced resting [Ca(2+)](i) in old mdx mice, suggesting that SACs may be involved in the Ca(2+)-handling abnormalities in these animals. This finding was supported by immunoblotting data, which demonstrated that old mdx mice had increased protein expression of canonical transient receptor potential channel 1, a likely candidate protein for SACs. SACs may play a role in the pathogenesis of the heart failure associated with DMD. Early in the disease process and before the onset of clinical symptoms increased, SAC activity may underlie the abnormal Ca(2+) handling in young mdx mice.
authors.affiliation: Bosch Institute, School of Medical Sciences, University of Sydney F13, NSW 2006 Australia.
authors.forename: Iwan A
authors.initials: IA
authors.lastname: Williams
authors.forename: David G
authors.initials: DG
authors.lastname: Allen
mesh_terms: D000367:Age Factors; D000818:Animals; D002110:Caffeine / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000378:metabolism; D002303:Cardiac Output, Low / Q000209:etiology / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology; D002311:Cardiomyopathy, Dilated / Q000209:etiology / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology; D004195:Disease Models, Animal; D004558:Electric Stimulation; D005355:Fibrosis; D006352:Heart Ventricles / Q000378:metabolism / Q000503:physiopathology; D036341:Intercellular Signaling Peptides and Proteins; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018101:Mice, Inbred mdx; D009137:Muscular Dystrophy, Animal / Q000150:complications; D020388:Muscular Dystrophy, Duchenne / Q000150:complications*; D009200:Myocardial Contraction; D032383:Myocytes, Cardiac / Q000187:drug effects / Q000378:metabolism*; D010455:Peptides / Q000494:pharmacology; D010766:Phosphorylation; D019837:Ryanodine Receptor Calcium Release Channel / Q000378:metabolism; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases / Q000378:metabolism; D019831:Sodium-Calcium Exchanger / Q000187:drug effects / Q000378:metabolism; D013111:Spider Venoms / Q000494:pharmacology; D013307:Streptomycin / Q000494:pharmacology; D050052:TRPC Cation Channels / Q000187:drug effects / Q000378:metabolism; D013997:Time Factors; D000099211:Phospholamban
chemical_list: D002110:Caffeine; D002118:Calcium; D002135:Calcium-Binding Proteins; D036341:Intercellular Signaling Peptides and Proteins; D010455:Peptides; D019837:Ryanodine Receptor Calcium Release Channel; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases; D019831:Sodium-Calcium Exchanger; D013111:Spider Venoms; D013307:Streptomycin; D050052:TRPC Cation Channels; D000099211:Phospholamban; C411249:MTx4 protein, Grammostola spatulata; C096351:transient receptor potential cation channel, subfamily C, member 1; C506947:Atp2a2 protein, mouse