BRIEF RESEARCH REPORT article
Front. Physiol.
Sec. Cardiac Electrophysiology
Volume 16 - 2025 | doi: 10.3389/fphys.2025.1579815
Simultaneous optical recording of action potentials and calcium transients in cardiac single cells differentiated from type 1 CPVT-iPS cells
Provisionally accepted
- 1Center for iPS Cell Research and Application, Kyoto University, Kyoto, Japan
- 2Takeda (Japan), Osaka, Japan
- 3Takeda Pharmaceutical Company Limited, Capital Federal, Argentina
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Numerous reports investigating channelopathies, including Catecholaminergic Polymorphic Ventricular Tachycardia (CPVT), have successfully reproduced using cardiomyocytes (CMs) differentiated from human induced pluripotent stem cells (hiPSCs). However, the relationship between action potentials (AP) and calcium transient waveforms-especially after drug treatment-remains unclear. In this study, we simultaneously loaded a membrane potential dye FluoVolt and the new calcium indicator Calbryte TM 590 AM and optimized stimulation and detection of both dyes to successfully obtain a higher signal-to-noise (S/N) ratio than the conventional membrane potential dye-red fluorescence Ca 2+ dye combination, thus enabling the simultaneous recording of both AP and calcium transient waveforms in single hiPSC-CMs, which continued even after gradual increases in drug concentration. In drug-loading experiments on CPVT1 (RyR2-I4587V) hiPSC-derived ventricular-like CMs, carvedilol and flecainide demonstrated some effectiveness, while JTV519 at 3µM exhibited both efficacy and alterations in AP waveforms. The Ca 2+ /calmodulin-dependent serine-threonine protein kinase II (CaMKII) inhibitor KN-93 at 1 µM was highly effective (93%) at reducing Ca 2+ transient abnormalities without altering AP waveforms.
Keywords: FluoVolt, Membrane potential dye, Calbryte 590, Calcium transient, Catecholaminergic polymorphic ventricular tachycardia, Induced Pluripotent Stem Cells, JTV519, KN-93
Received: 19 Feb 2025; Accepted: 12 May 2025.
Copyright: © 2025 Takaki, Tamura, Imahashi, Nishimoto and Yoshida. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Yoshinori Yoshida, Center for iPS Cell Research and Application, Kyoto University, Kyoto, Japan
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