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Circulation Research. 1999;84:266-275

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(Circulation Research. 1999;84:266-275.)
© 1999 American Heart Association, Inc.


Original Contribution

Examination of the Transverse Tubular System in Living Cardiac Rat Myocytes by 2-Photon Microscopy and Digital Image–Processing Techniques

C. Soeller, M. B. Cannell

From the Department of Physiology, University of Auckland, Auckland, NZ.

Correspondence to Dr Mark Cannell, Department of Physiology, University of Auckland, Private Bag 92019, Grafton St, Auckland, NZ. E-mail m.cannell{at}auckland.ac.nz

Abstract—The transverse tubular system (t-system) of cardiac muscle is a structure that allows rapid propagation of excitation into the cell interior. Using 2-photon molecular excitation microscopy and digital image–processing methods, we have obtained a comprehensive overview of the t-system of rat ventricular myocytes in living cells. We show that it is possible to quantify the morphology of the t-system in terms of average local tubule diameter, branching pattern, and local abundance of the t-system by immersing living myocytes in a dextran-linked fluorescein solution. Our data suggest that previous electron microscopic examinations of t-system structure have underestimated both the geometric complexity of the t-system morphology and the fraction of cell volume occupied by the t-system (3.6% in this species). About 40% of tubules occur between Z-lines, and the t-tubule diameter is 255±0.85 nm (mean±SEM). The t-tubules leave the outer surface of the cell in an approximately rectangular array; however, at some points junctions between the t-tubules and the surface membrane are missing. In view of the complexity of the t-system apparent from our images, we propose that the t-system be renamed the "sarcolemmal Z rete." The methods presented here are generally applicable to the quantification of the sarcolemmal Z rete and other structures within cells by fluorescence microscopy in a variety of cell types.


Key Words: heart structure • t-system • imaging • myocyte • rat




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