changes in the organization of excitation-contraction coupling structures in failing human heart组织的变化没有人类心脏兴奋收缩偶联结构.pdfVIP

Changes in the Organization of Excitation-Contraction Coupling Structures in Failing Human Heart 1 1,2 1 1 David J. Crossman , Peter R. Ruygrok , Christian Soeller , Mark B. Cannell * 1 Department of Physiology, Faculty of Medicine and Health Sciences, University of Auckland, Auckland, New Zealand, 2 Auckland City Hospital, Auckland, New Zealand Abstract Background: The cardiac myocyte t-tubular system ensures rapid, uniform cell activation and several experimental lines of evidence suggest changes in the t-tubular system and associated excitation-contraction coupling proteins may occur in heart failure. Methods and Results: The organization of t-tubules, L-type calcium channels (DHPRs), ryanodine receptors (RyRs) and contractile machinery were examined in fixed ventricular tissue samples from both normal and failing hearts (idiopathic (non-ischemic) dilated cardiomyopathy) using high resolution fluorescent imaging. Wheat germ agglutinin (WGA), Na-Ca exchanger, DHPR and caveolin-3 labels revealed a shift from a predominantly transverse orientation to oblique and axial directions in failing myocytes. In failure, dilation of peripheral t-tubules occurred and a change in the extent of protein glycosylation was evident. There was no change in the fractional area occupied by myofilaments (labeled with phalloidin) but there was a small reduction in the number of RyR clusters per unit area. The general relationship between DHPRs and RyR was not changed and RyR labeling overlapped with 5163% of DHPR labeling in normal hearts. In longitudinal (but not transverse) sections there was an ,30% reduction in the degree of colocalization between DHPRs and RyRs as measured by Pearson’s correlation coefficient in fa