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Comparing the effects of chemical Ca2+ dyes and R-GECO on contractility and Ca2+ transients in adult and human iPSC cardiomyocytes

Robinson, Paul, Sparrow, Alexander J., Psaras, Yiangos, Steeples, Violetta, Simon, Jillian, Broyles, Connor N., Chang, Yu-Fen, Brook, Frances A., Wang, Ying-Jie, Blease, Andrew, and others. (2023) Comparing the effects of chemical Ca2+ dyes and R-GECO on contractility and Ca2+ transients in adult and human iPSC cardiomyocytes. Journal of Molecular and Cellular Cardiology, 180 . pp. 44-57. ISSN 0022-2828. E-ISSN 1095-8584. (doi:10.1016/j.yjmcc.2023.04.008) (KAR id:101223)

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Abstract

We compared commonly used BAPTA-derived chemical Ca2+ dyes (fura2, Fluo-4, and Rhod-2) with a newer genetically encoded indicator (R-GECO) in single cell models of the heart. We assessed their performance and effects on cardiomyocyte contractility, determining fluorescent signal-to-noise ratios and sarcomere shortening in primary ventricular myocytes from adult mouse and guinea pig, and in human iPSC-derived cardiomyocytes. Chemical Ca2+ dyes displayed dose-dependent contractile impairment in all cell types, and we observed a negative correlation between contraction and fluorescence signal-to-noise ratio, particularly for fura2 and Fluo-4. R-GECO had no effect on sarcomere shortening. BAPTA-based dyes, but not R-GECO, inhibited in vitro acto-myosin ATPase activity. The presence of fura2 accentuated or diminished changes in contractility and Ca2+ handling caused by small molecule modulators of contractility and intracellular ionic homeostasis (mavacamten, levosimendan, and flecainide), but this was not observed when using R-GECO in adult guinea pig left ventricular cardiomyocytes. Ca2+ handling studies are necessary for cardiotoxicity assessments of small molecules intended for clinical use. Caution should be exercised when interpreting small molecule studies assessing contractile effects and Ca2+ transients derived from BAPTA-like chemical Ca2+ dyes in cellular assays, a common platform for cardiac toxicology testing and mechanistic investigation of cardiac disease physiology and treatment.

Item Type: Article
DOI/Identification number: 10.1016/j.yjmcc.2023.04.008
Subjects: Q Science > Q Science (General)
Divisions: Divisions > Division of Natural Sciences > Biosciences
Funders: University of Kent (https://ror.org/00xkeyj56)
Depositing User: Michael Geeves
Date Deposited: 11 May 2023 08:51 UTC
Last Modified: 04 Jul 2023 13:36 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/101223 (The current URI for this page, for reference purposes)

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