Abstract
The possibility to generate cardiomyocytes (CMs) from disease-specific induced pluripotent stem cells (iPSCs) is a powerful tool for the investigation of various cardiac diseases in vitro. The pathological course of various cardiac conditions, causatively heterogeneous, often converges into disturbed cellular Ca(2+) cycling. The gigantic Ca(2+) channel of the intracellular Ca(2+) store of CMs, the ryanodine receptor type 2 (RyR2), controls Ca(2+) release and therefore plays a crucial role in Ca(2+) cycling of CMs. In the present protocol we describe ways to measure and analyze global as well as local cellular Ca(2+) release events in CMs derived from a patient carrying a CPVT-causing RyR2 mutation.
Keywords:
Calcium transients; Cardiomyocytes; Confocal microscopy; Sparks; Tachycardia.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Aniline Compounds
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Animals
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Calcium / metabolism
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Calcium Signaling*
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Cell Culture Techniques / methods*
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Cell Differentiation
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Cellular Reprogramming*
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Fibroblasts / cytology
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Fibroblasts / metabolism*
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Gene Expression
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Humans
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Induced Pluripotent Stem Cells / drug effects
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Induced Pluripotent Stem Cells / metabolism*
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Induced Pluripotent Stem Cells / ultrastructure
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Isoproterenol / pharmacology
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Mice
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Molecular Imaging / methods
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Mutation
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Myocytes, Cardiac / drug effects
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Myocytes, Cardiac / metabolism*
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Myocytes, Cardiac / ultrastructure
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Primary Cell Culture
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Ryanodine / pharmacology
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Ryanodine Receptor Calcium Release Channel / genetics
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Ryanodine Receptor Calcium Release Channel / metabolism
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Tachycardia / genetics
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Tachycardia / metabolism
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Tachycardia / pathology
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Xanthenes
Substances
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Aniline Compounds
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Fluo 4
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Ryanodine Receptor Calcium Release Channel
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Xanthenes
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Ryanodine
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Isoproterenol
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Calcium