Key result
A systematic approach using fluorescent Ca2+ indicators and whole-cell recording allows detailed evaluation of essential transport mechanisms and Ca2+ fluxes in isolated single cardiac myocytes.
This review outlines a systematic methodological approach for assessing calcium handling and ryanodine receptor function in isolated cardiac myocytes.
Standardizes Ca2+ flux assessment in isolated myocytes; extends research methodology but leaves clinical translation open pending validation.
In cardiac myocytes, Ca(2+) release from the sarcoplasmic reticulum (SR) Ca(2+) store through the opening of ryanodine receptors (RyRs) is the major source of Ca(2+) for activation of myofilaments and contraction. Over the past 20 years, tools have become available to study this release process in detail, allowing new insights into the regulation of SR Ca(2+) release and RyR function. To assess these processes, we recommend and here review a systematic approach that evaluates the essential transport mechanisms and Ca(2+) fluxes in isolated single cardiac myocytes by using fluorescent Ca(2+) indicators and whole-cell recording of membrane voltage and ionic currents under voltage clamp. The approach includes an assessment of the L-type Ca(2+) current as a trigger for opening of RyRs and release of SR Ca(2+), of the SR Ca(2+) content, of intrinsic properties of RyRs, and of Ca(2+)-removal systems.
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Sipido et al. (2015) conducted a review in Cardiac myocyte Ca2+ handling. Systematic approach for assessing Ca2+ handling was evaluated. A systematic approach using fluorescent Ca2+ indicators and whole-cell recording allows detailed evaluation of essential transport mechanisms and Ca2+ fluxes in isolated single cardiac myocytes.
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