Measuring Ca²⁺ sparks in cardiac myocytes

MacQuaide, N. , Bito, V. and Sipido, K. R. (2015) Measuring Ca²⁺ sparks in cardiac myocytes. Cold Spring Harbor Protocols, 2015(5), pp. 490-7. (doi: 10.1101/pdb.prot076984) (PMID:25934930)

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Abstract

This protocol describes the measurement of Ca(2+) sparks in intact myocytes by using a Ca(2+)-sensitive dye and imaging using laser scanning confocal microscopy. It takes advantage of spontaneous Ca(2+)-release events-sparks-using them as a measure of the activity of ryanodine receptors (RyRs). Two methodologies are described: One requires that cardiomyocytes be stimulated, preferably under voltage clamp by depolarizing pulses, until steady-state is reached, and then stimulation is stopped and Ca(2+) sparks are recorded. The second requires that cells be permeabilized and bathed in a solution to load the cell with Ca(2+) sufficient to elicit Ca(2+) sparks, but not Ca(2+) waves. These are then analyzed offline to quantify spark frequency and morphology. The advantages and disadvantages of each approach are discussed.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:MacQuaide, Dr Niall
Authors: MacQuaide, N., Bito, V., and Sipido, K. R.
College/School:College of Medical Veterinary and Life Sciences > Institute of Cardiovascular and Medical Sciences
Journal Name:Cold Spring Harbor Protocols
Publisher:Cold Spring Harbor Laboratory Press
ISSN:1940-3402
ISSN (Online):1559-6095

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