NDLI: Measuring $Ca^{2+}$ Sparks in Cardiac Myocytes

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Measuring $Ca^{2+}$ Sparks in Cardiac Myocytes

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Macquaide, Niall Bito, Virginie Sipido, Karin R.
Description	Country affiliation: Belgium Author Affiliation: Macquaide N ( Division of Experimental Cardiology, Department of Cardiovascular Sciences, KU Leuven, Belgium.); Bito V ( Division of Experimental Cardiology, Department of Cardiovascular Sciences, KU Leuven, Belgium.); Sipido KR ( Division of Experimental Cardiology, Department of Cardiovascular Sciences, KU Leuven, Belgium.)
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.
File Format	HTM / HTML
ISSN	19403402
Issue Number	5
Volume Number	2015
e-ISSN	15596095
Journal	Cold Spring Harbor Protocols
Language	English
Publisher	Cold Spring Harbor Laboratory Press
Publisher Date	2015-05-01
Publisher Place	United States
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Discipline Clinical Laboratory Techniques Calcium Analysis Cytological Techniques Methods Fluorescent Dyes Metabolism Microscopy, Confocal Myocytes, Cardiac Chemistry Patch-clamp Techniques Staining And Labeling Journal Article Research Support, Non-u.s. Gov't
Content Type	Text
Resource Type	Article
Subject	Biochemistry, Genetics and Molecular Biology

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A Systematic Approach for Assessing $Ca^{2+}$ Handling in Cardiac Myocytes

Measuring Sarcoplasmic Reticulum $Ca^{2+}$ Content, Fractional Release, and $Ca^{2+}$ Buffering in Cardiac Myocytes

Imaging of Mitochondrial and Nuclear $Ca^{2+}$ Dynamics in Arabidopsis Roots

The quantal nature of $Ca^{2+}$ sparks and in situ operation of the ryanodine receptor array in cardiac cells

Basic Methods for Monitoring Intracellular $Ca^{2+}$ in Cardiac Myocytes Using Fluo-3

$Ca^{2+}$ Imaging in Plants Using Genetically Encoded Yellow Cameleon $Ca^{2+}$ Indicators

Live Cell Imaging of Cytoplasmic and Nuclear $Ca^{2+}$ Dynamics in Arabidopsis Roots

Fluorescent speckle microscopy.

Measurement of Free $Ca^{2+}$ Concentration in the Lumen of Neuronal Endoplasmic Reticulum

Measuring $Ca^{2+}$ Sparks in Cardiac Myocytes

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A Systematic Approach for Assessing $Ca^{2+}$ Handling in Cardiac Myocytes

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