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Open AccessArticle

Mapping Calcium Dynamics in the Heart of Zebrafish Embryos with Ratiometric Genetically Encoded Calcium Indicators

1
Physiology and Cell Dynamics, Centro Regional de Investigaciones Biomédicas (CRIB), Facultad de Medicina de Albacete, Departamento de Ciencias Médicas, Universidad de Castilla-La Mancha, C/Almansa 14, 02006 Albacete, Spain
2
UMR8256, Biological Adaptation and Ageing, CNRS, Sorbonne Université, F-75005 Paris, France
*
Authors to whom correspondence should be addressed.
Int. J. Mol. Sci. 2020, 21(18), 6610; https://doi.org/10.3390/ijms21186610
Received: 4 September 2020 / Accepted: 8 September 2020 / Published: 10 September 2020
Zebrafish embryos have been proposed as a cost-effective vertebrate model to study heart function. Many fluorescent genetically encoded Ca2+ indicators (GECIs) have been developed, but those with ratiometric readout seem more appropriate to image a moving organ such as the heart. Four ratiometric GECIs based on troponin C, TN-XXL, Twitch-1, Twitch-2B, and Twitch-4 were expressed transiently in the heart of zebrafish embryos. Their emission ratio reported the Ca2+ levels in both the atrium and the ventricle. We measured several kinetic parameters of the Ca2+ transients: systolic and diastolic ratio, the amplitude of the systolic Ca2+ rise, the heart rate, as well as the rise and decay times and slopes. The systolic ratio change decreased in cells expressing high biosensor concentration, possibly caused by Ca2+ buffering. The GECIs were able to report the effect of nifedipine and propranolol on the heart, which resulted in changes in heart rate, diastolic and systolic Ca2+ levels, and Ca2+ kinetics. As a result, Twitch-1 and Twitch-4 (Kd 0.25 and 2.8 µM, respectively) seem the most promising GECIs for generating transgenic zebrafish lines, which could be used for modeling heart disorders, for drug screening, and for cardiotoxicity assessment during drug development. View Full-Text
Keywords: calcium; genetically encoded calcium indicator (GECI); biosensor; zebrafish; embryo; heart; imaging calcium; genetically encoded calcium indicator (GECI); biosensor; zebrafish; embryo; heart; imaging
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MDPI and ACS Style

Salgado-Almario, J.; Vicente, M.; Vincent, P.; Domingo, B.; Llopis, J. Mapping Calcium Dynamics in the Heart of Zebrafish Embryos with Ratiometric Genetically Encoded Calcium Indicators. Int. J. Mol. Sci. 2020, 21, 6610. https://doi.org/10.3390/ijms21186610

AMA Style

Salgado-Almario J, Vicente M, Vincent P, Domingo B, Llopis J. Mapping Calcium Dynamics in the Heart of Zebrafish Embryos with Ratiometric Genetically Encoded Calcium Indicators. International Journal of Molecular Sciences. 2020; 21(18):6610. https://doi.org/10.3390/ijms21186610

Chicago/Turabian Style

Salgado-Almario, Jussep; Vicente, Manuel; Vincent, Pierre; Domingo, Beatriz; Llopis, Juan. 2020. "Mapping Calcium Dynamics in the Heart of Zebrafish Embryos with Ratiometric Genetically Encoded Calcium Indicators" Int. J. Mol. Sci. 21, no. 18: 6610. https://doi.org/10.3390/ijms21186610

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