Simultaneous imaging of calcium and contraction in the beating heart of zebrafish larvae.

In vivo models of cardiac function maintain the complex relationship of cardiomyocytes with other heart cells,as well as the paracrine and mechanoelectrical feedback mechanisms. We aimed at imaging calcium transients simultaneously with heart contraction in zebrafish larvae. Methods: To image calciu...

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Autores: Salgado Almario, Jussep Alfredo, Vicente Ruiz, Manuel, Molina Durango, Yillcer, Martínez Sielva, Antonio, Vincent , Pierre, Domingo Moreno, Beatriz, Llopis Borrás, Juan Francisco
Tipo de recurso: artículo
Fecha de publicación:2022
País:España
Institución:Universidad de Castilla-La Mancha
Repositorio:RUIdeRA. Repositorio Institucional de la UCLM
OAI Identifier:oai:ruidera.uclm.es:10578/39892
Acceso en línea:https://doi.org/10.7150/thno.64734
https://hdl.handle.net/10578/39892
Access Level:acceso abierto
Palabra clave:Calcium
Contractation
Heart
Imaging
Zebrafish
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spelling Simultaneous imaging of calcium and contraction in the beating heart of zebrafish larvae.Salgado Almario, Jussep AlfredoVicente Ruiz, ManuelMolina Durango, YillcerMartínez Sielva, AntonioVincent , PierreDomingo Moreno, BeatrizLlopis Borrás, Juan FranciscoCalciumContractationHeartImagingZebrafishIn vivo models of cardiac function maintain the complex relationship of cardiomyocytes with other heart cells,as well as the paracrine and mechanoelectrical feedback mechanisms. We aimed at imaging calcium transients simultaneously with heart contraction in zebrafish larvae. Methods: To image calcium in beating hearts, we generated a zebrafish transgenic line expressing the FRET-based ratiometric biosensor Twitch-4. Since emission ratioing canceled out the motion artifacts, we did not use myosin inhibitors or tnnt2a morpholinos to uncouple contraction from changes in calcium levels. We wrote an analysis program to automatically calculate kinetic parameters of the calcium transients. In addition, the ventricular diameter was determined in the fluorescence images providing a real-time measurement of contraction correlated with calcium. Results: Expression of Twitch-4 did not affect the force of contraction, the size of the heart nor the heart rate in 3- and 5-days post-fertilization (dpf) larvae. Comparison of 3 and 5 dpf larvae showed that calcium levels and transient amplitude were larger at 5 dpf, but the fractional shortening did not change. To validate the model, we evaluated the effect of drugs with known effects on cardiomyocytes. Calcium levels and the force of contraction decreased by the L-type calcium channel blocker nifedipine, whereas they increased with the activator Bay-K 8644. Caffeine induced bradycardia, markedly decreased ventricular diastolic calcium levels, increased the size of the calcium transients, and caused an escape rhythm in some larvae. Conclusions: The Tg(myl7:Twitch-4) line provides a physiological approach to image systolic and diastolic calcium levels in the heart of zebrafish larvae. Since the heart is beating, calcium levels and contraction can be correlated. This line will be a useful tool to address pathophysiological mechanisms in diseases like heart failure and arrhythmia, in cardiotoxicity studies and for drug screening.Ivyspring202420242022info:eu-repo/semantics/articleapplication/pdfapplication/pdfhttps://doi.org/10.7150/thno.64734https://hdl.handle.net/10578/39892reponame:RUIdeRA. Repositorio Institucional de la UCLMinstname:Universidad de Castilla-La ManchaInglésPID2019-111456RB-100BFU2015-69874-REQC-2019-006102-PSBPLY/19/180501/0002232019-GRIN-270192020-GRIN-291862021-GRIN-31151info:eu-repo/semantics/openAccessoai:ruidera.uclm.es:10578/398922026-05-27T07:36:41Z
dc.title.none.fl_str_mv Simultaneous imaging of calcium and contraction in the beating heart of zebrafish larvae.
title Simultaneous imaging of calcium and contraction in the beating heart of zebrafish larvae.
spellingShingle Simultaneous imaging of calcium and contraction in the beating heart of zebrafish larvae.
Salgado Almario, Jussep Alfredo
Calcium
Contractation
Heart
Imaging
Zebrafish
title_short Simultaneous imaging of calcium and contraction in the beating heart of zebrafish larvae.
title_full Simultaneous imaging of calcium and contraction in the beating heart of zebrafish larvae.
title_fullStr Simultaneous imaging of calcium and contraction in the beating heart of zebrafish larvae.
title_full_unstemmed Simultaneous imaging of calcium and contraction in the beating heart of zebrafish larvae.
title_sort Simultaneous imaging of calcium and contraction in the beating heart of zebrafish larvae.
dc.creator.none.fl_str_mv Salgado Almario, Jussep Alfredo
Vicente Ruiz, Manuel
Molina Durango, Yillcer
Martínez Sielva, Antonio
Vincent , Pierre
Domingo Moreno, Beatriz
Llopis Borrás, Juan Francisco
author Salgado Almario, Jussep Alfredo
author_facet Salgado Almario, Jussep Alfredo
Vicente Ruiz, Manuel
Molina Durango, Yillcer
Martínez Sielva, Antonio
Vincent , Pierre
Domingo Moreno, Beatriz
Llopis Borrás, Juan Francisco
author_role author
author2 Vicente Ruiz, Manuel
Molina Durango, Yillcer
Martínez Sielva, Antonio
Vincent , Pierre
Domingo Moreno, Beatriz
Llopis Borrás, Juan Francisco
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv Calcium
Contractation
Heart
Imaging
Zebrafish
topic Calcium
Contractation
Heart
Imaging
Zebrafish
description In vivo models of cardiac function maintain the complex relationship of cardiomyocytes with other heart cells,as well as the paracrine and mechanoelectrical feedback mechanisms. We aimed at imaging calcium transients simultaneously with heart contraction in zebrafish larvae. Methods: To image calcium in beating hearts, we generated a zebrafish transgenic line expressing the FRET-based ratiometric biosensor Twitch-4. Since emission ratioing canceled out the motion artifacts, we did not use myosin inhibitors or tnnt2a morpholinos to uncouple contraction from changes in calcium levels. We wrote an analysis program to automatically calculate kinetic parameters of the calcium transients. In addition, the ventricular diameter was determined in the fluorescence images providing a real-time measurement of contraction correlated with calcium. Results: Expression of Twitch-4 did not affect the force of contraction, the size of the heart nor the heart rate in 3- and 5-days post-fertilization (dpf) larvae. Comparison of 3 and 5 dpf larvae showed that calcium levels and transient amplitude were larger at 5 dpf, but the fractional shortening did not change. To validate the model, we evaluated the effect of drugs with known effects on cardiomyocytes. Calcium levels and the force of contraction decreased by the L-type calcium channel blocker nifedipine, whereas they increased with the activator Bay-K 8644. Caffeine induced bradycardia, markedly decreased ventricular diastolic calcium levels, increased the size of the calcium transients, and caused an escape rhythm in some larvae. Conclusions: The Tg(myl7:Twitch-4) line provides a physiological approach to image systolic and diastolic calcium levels in the heart of zebrafish larvae. Since the heart is beating, calcium levels and contraction can be correlated. This line will be a useful tool to address pathophysiological mechanisms in diseases like heart failure and arrhythmia, in cardiotoxicity studies and for drug screening.
publishDate 2022
dc.date.none.fl_str_mv 2022
2024
2024
dc.type.none.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv https://doi.org/10.7150/thno.64734
https://hdl.handle.net/10578/39892
url https://doi.org/10.7150/thno.64734
https://hdl.handle.net/10578/39892
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv PID2019-111456RB-100
BFU2015-69874-R
EQC-2019-006102-P
SBPLY/19/180501/000223
2019-GRIN-27019
2020-GRIN-29186
2021-GRIN-31151
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Ivyspring
publisher.none.fl_str_mv Ivyspring
dc.source.none.fl_str_mv reponame:RUIdeRA. Repositorio Institucional de la UCLM
instname:Universidad de Castilla-La Mancha
instname_str Universidad de Castilla-La Mancha
reponame_str RUIdeRA. Repositorio Institucional de la UCLM
collection RUIdeRA. Repositorio Institucional de la UCLM
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,812429

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