title: Ultrastructural and functional alterations of EC coupling elements in mdx cardiomyocytes: an analysis from membrane surface to depth.
abstract: A dilated cardiomyopathy (DCM) is associated with Duchenne muscular dystrophy (DMD). The loss of dystrophin leads to membrane instability and calcium dysregulation in skeletal muscle but effects of such a loss are not elucidated at cardiomyocytes level. We sought to examine whether membrane and transverse tubules damages occur in ventricular myocytes from mdx mouse model of DMD and how they impact the function of single excitation-contraction coupling elements. Scanning ion conductance microscopy (SICM) was used to characterize the integrity loss of living mdx cardiomyocytes surface. 2D Fourier transform analysis of labeled internal networks (transverse tubules, alpha-actinin, dihydropyridine receptors, ryanodine receptors) was performed to evaluate internal alterations. During calcium measurements, "smart microperfusions" of depolarizing solutions were applied through SICM nanopipette, stimulating single tubules elements. These approaches revealed structural membrane surface (39% decrease for Z-groove ratio) and transverse tubules disorganization (21% transverse tubules ratio decrease) in mdx as compared to control. These disruptions were associated with functional alterations (sixfold increase of calcium signal duration and twofold increase of sparks frequency). In DCM associated with DMD, myocytes display evident membrane alterations at the surface level but also in the cell depth with a disruption of transverse tubules network as observed in other cases of heart failure. These ultrastructural changes are associated with changes in the function of some coupling elements. Thus, these profound disruptions may play a role in calcium dysregulation through excitation-contraction coupling elements perturbation and suggest a transverse tubules stabilizing role for dystrophin.
authors: Institut de Physiologie et Biologie Cellulaires, FRE CNRS/Université de Poitiers n°3511, Pôle Biologie Santé Bât B36, BP 633, Poitiers, France.
authors: Charlotte
authors: C
authors: Lorin
authors: Mélanie
authors: M
authors: Gueffier
authors: Patrick
authors: P
authors: Bois
authors: Jean-François
authors: JF
authors: Faivre
authors: Christian
authors: C
authors: Cognard
authors: Stéphane
authors: S
authors: Sebille
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism; D002311:Cardiomyopathy, Dilated / Q000473:pathology; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure*; D056966:Excitation Contraction Coupling; D042541:Intracellular Space / Q000378:metabolism; D008297:Male; D051379:Mice; D018101:Mice, Inbred mdx; D057054:Molecular Imaging; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000648:ultrastructure; D012508:Sarcolemma / Q000378:metabolism / Q000648:ultrastructure
chemical_list: D002118:Calcium