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)

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 > School of Cardiovascular & Metabolic Health
• Journal Name: Cold Spring Harbor Protocols
• Publisher: Cold Spring Harbor Laboratory Press
• ISSN: 1940-3402
• ISSN (Online): 1559-6095