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Am J Physiol Heart Circ Physiol 278: H666-H669, 2000;
0363-6135/00 $5.00
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Vol. 278, Issue 2, H666-H669, February 2000

RAPID COMMUNICATION
Caffeine-induced Ca2+ sparks in mouse ventricular myocytes

Michael Ritter1, Zhi Su1, Kenneth W. Spitzer2, Hideyuki Ishida3, and William H. Barry1

1 Division of Cardiology and 2 Nora Eccles Harrison Cardiovascular Research and Training Institute, University of Utah Health Sciences Center, Salt Lake City, Utah 84132; and 3 Department of Physiology, Tokai University, Kanagawa 259-1193, Japan

Ca2+ sparks are spatially localized intracellular Ca2+ release events that were first described in 1993. Sparks have been ascribed to sarcoplasmic reticulum Ca2+ release channel (ryanodine receptor, RyR) opening induced by Ca2+ influx via L-type Ca2+ channels or by spontaneous RyR openings and have been thought to reflect Ca2+ release from a cluster of RyR. Here we describe a pharmacological approach to study sparks by exposing ventricular myocytes to caffeine with a rapid solution-switcher device. Sparks under these conditions have properties similar to naturally occurring sparks in terms of size and intracellular Ca2+ concentration ([Ca2+]i) amplitude. However, after the diffusion of caffeine, sparks first appear close to the cell surface membrane before coalescing to produce a whole cell transient. Our results support the idea that a whole cell [Ca2+]i transient consists of the summation of sparks and that Ca2+ sparks consist of the opening of a cluster of RyR and confirm that characteristics of the cluster rather than the L-type Ca2+ channel-RyR relation determine spark properties.

ryanodine receptors; confocal microscopy


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