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Cardiology Research and Practice
Volume 2009 (2009), Article ID 802373, 9 pages
http://dx.doi.org/10.4061/2009/802373
Research Article

Two-Photon Laser Scanning Microscopy of the Transverse-Axial Tubule System in Ventricular Cardiomyocytes from Failing and Non-Failing Human Hearts

1Department of Medicine, Institute of Molecular Cardiobiology, Johns Hopkins University, Baltimore, MD 21205, USA
2Department of Cardiology and Pneumology, Georg-August-University Göttingen, 37075 Göttingen, Germany
3Department of Physiology, Cardiovascular Research Center, Temple University School of Medicine, Philadelphia, PA 19140, USA
4Department of Cardiology, University of Bonn, 53105 Bonn, Germany
5Department of Medicine, University School of Medicine, Durham, NC 27705, USA

Received 7 August 2009; Accepted 17 November 2009

Academic Editor: Chim C. Lang

Copyright © 2009 Andreas Ohler et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Objective. The transverse-axial tubule system (TATS) of cardiomyocytes allows a spatially coordinated conversion of electrical excitation into an intracellular signal and consequently contraction. Previous reports have indicated alterations of structure and/or volume of the TATS in cardiac hypertrophy and failure, suggesting a contribution to the impairment of excitation contraction coupling. To test whether structural alterations are present in human heart failure, the TATS was visualized in myocytes from failing and non-failing human hearts. Methods and Results. In freshly isolated myocytes, the plasmalemmal membranes were labeled with Di-8-ANEPPS and imaged using two-photon excitation at 780 nm. Optical sections were taken every 300 nm through the cells. After deconvolution, the TATS was determined within the 3D data sets, revealing no significant difference in normalized surface area or volume. To rule out possible inhomogeneity in the arrangement of the TATS, Euclidian distance maps were plotted for every section, allowing to measure the closest distance between any cytosolic and any membrane point. There was a trend towards greater spacing in cells from failing hearts, without statistical significance. Conclusion. Only small changes, but no significant changes in the geometrical dimensions of the TATS were observed in cardiomyocytes from failing compared to non-failing human myocardium.