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01-07-2017 | Original Contribution

The role of spatial organization of Ca²⁺ release sites in the generation of arrhythmogenic diastolic Ca²⁺ release in myocytes from failing hearts

Authors: Andriy E. Belevych, Hsiang-Ting Ho, Ingrid M. Bonilla, Radmila Terentyeva, Karsten E. Schober, Dmitry Terentyev, Cynthia A. Carnes, Sándor Györke

Published in: Basic Research in Cardiology | Issue 4/2017

Abstract

In heart failure (HF), dysregulated cardiac ryanodine receptors (RyR2) contribute to the generation of diastolic Ca²⁺ waves (DCWs), thereby predisposing adrenergically stressed failing hearts to life-threatening arrhythmias. However, the specific cellular, subcellular, and molecular defects that account for cardiac arrhythmia in HF remain to be elucidated. Patch-clamp techniques and confocal Ca²⁺ imaging were applied to study spatially defined Ca²⁺ handling in ventricular myocytes isolated from normal (control) and failing canine hearts. Based on their activation time upon electrical stimulation, Ca²⁺ release sites were categorized as coupled, located in close proximity to the sarcolemmal Ca²⁺ channels, and uncoupled, the Ca²⁺ channel-free non-junctional Ca²⁺ release units. In control myocytes, stimulation of β-adrenergic receptors with isoproterenol (Iso) resulted in a preferential increase in Ca²⁺ spark rate at uncoupled sites. This site-specific effect of Iso was eliminated by the phosphatase inhibitor okadaic acid, which caused similar facilitation of Ca²⁺ sparks at coupled and uncoupled sites. Iso-challenged HF myocytes exhibited increased predisposition to DCWs compared to control myocytes. In addition, the overall frequency of Ca²⁺ sparks was increased in HF cells due to preferential stimulation of coupled sites. Furthermore, coupled sites exhibited accelerated recovery from functional refractoriness in HF myocytes compared to control myocytes. Spatially resolved subcellular Ca²⁺ mapping revealed that DCWs predominantly originated from coupled sites. Inhibition of CaMKII suppressed DCWs and prevented preferential stimulation of coupled sites in Iso-challenged HF myocytes. These results suggest that CaMKII- (and phosphatase)-dependent dysregulation of junctional Ca²⁺ release sites contributes to Ca²⁺-dependent arrhythmogenesis in HF.

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Title: The role of spatial organization of Ca2+ release sites in the generation of arrhythmogenic diastolic Ca2+ release in myocytes from failing hearts
Authors: Andriy E. Belevych
Hsiang-Ting Ho
Ingrid M. Bonilla
Radmila Terentyeva
Karsten E. Schober
Dmitry Terentyev
Cynthia A. Carnes
Sándor Györke
Publication date: 01-07-2017
Publisher: Springer Berlin Heidelberg
Published in: Basic Research in Cardiology / Issue 4/2017
Print ISSN: 0300-8428
Electronic ISSN: 1435-1803
DOI: https://doi.org/10.1007/s00395-017-0633-2

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The role of spatial organization of Ca²⁺ release sites in the generation of arrhythmogenic diastolic Ca²⁺ release in myocytes from failing hearts

Abstract

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Abstract

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