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J Mol Cell Cardiol. 2013 May;58:32-40. doi: 10.1016/j.yjmcc.2012.11.004. Epub 2012 Nov 13.

Super-resolution imaging of EC coupling protein distribution in the heart.

Author information

1
Department of Physiology, School of Medical Sciences, University of Auckland, New Zealand. c.soeller@auckland.ac.nz

Abstract

The cardiac ryanodine receptor (RyR) plays a central role in the control of contractile function of the heart. In cardiac ventricular myocytes RyRs and associated Ca(2+) handling proteins, including membrane Ca(2+) channels, Ca(2+) pumps and other sarcolemmal and sarcoplasmic reticulum proteins interact to set the time course and amplitude of the electrically triggered cytosolic Ca(2+) transient. It has become increasingly clear that protein distribution and clustering on the nanometer scale is critical in determining the interaction of these proteins and the resulting properties of cardiac Ca(2+) handling. Such intricate near-molecular scale detail cannot be visualized with conventional fluorescence microscopy techniques (e.g. confocal microscopy) but it has recently become accessible with optical super-resolution techniques. These techniques retain the advantages of fluorescent marker technology, i.e. high specificity and excellent contrast, but have a spatial resolution approaching 10nm, i.e. objects not much further apart than 10nm can be distinguished, previously only attainable with electron microscopy. We review the use of these novel imaging techniques for the study of protein distribution in cardiac ventricular myocytes and discuss technical considerations as well as recent findings using super-resolution imaging. An emphasis is on single molecule localization based super-resolution approaches and their use to reveal the complexity of RyR cluster morphology, placement and relationship to other excitation-contraction coupling proteins. Super-resolution imaging approaches have already demonstrated their utility for the study of cardiac structure-function relationships and we anticipate that their use will rapidly increase and help improve our understanding of cardiac Ca(2+) regulation.

PMID:
23159441
DOI:
10.1016/j.yjmcc.2012.11.004
[Indexed for MEDLINE]

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