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Laser-targeted ablation of the zebrafish embryonic ventricle: A novel model of cardiac injury and repair

Matrone, Gianfranco; Taylor, Jonathan M.; Wilson, Kathryn S.; Baily, James; Love, Gordon D.; Girkin, John M.; Mullins, John J.; Tucker, Carl S.; Denvir, Martin A.

Laser-targeted ablation of the zebrafish embryonic ventricle: A novel model of cardiac injury and repair Thumbnail


Authors

Gianfranco Matrone

Jonathan M. Taylor

Kathryn S. Wilson

James Baily

Gordon D. Love

John J. Mullins

Carl S. Tucker

Martin A. Denvir



Abstract

Background: While the adult zebrafish (Danio rerio) heart demonstrates a remarkable capacity for self-renewal following apical resection little is known about the response to injury in the embryonic heart. Methods: Injury to the beating zebrafish embryo heart was induced by laser using a transgenic zebrafish expressing cardiomyocyte specific green fluorescent protein. Changes in ejection fraction (EF), heart rate (HR), and caudal vein blood flow (CVBF) assessed by video capture techniques were assessed at 2, 24 and 48 h post-laser. Change in total and mitotic ventricular cardiomyocyte number following laser injury was also assessed by counting respectively DAPI (VCt) and Phospho-histone H3 (VCm) positive nuclei in isolated hearts using confocal microscopy. Results: Laser injury to the ventricle resulted in bradycardia and mild bleeding into the pericardium. At 2 h post-laser injury, there was a significant reduction in cardiac performance in lasered-hearts compared with controls (HR 117 ± 11 vs 167 ± 9 bpm, p ≤ 0.001; EF 14.1 ± 1.8 vs 20.1 ± 1.3%, p ≤ 0.001; CVBF 103 ± 15 vs 316 ± 13μms− 1, p ≤ 0.001, respectively). Isolated hearts showed a significant reduction in VCt at 2 h post-laser compared to controls (195 ± 15 vs 238 ± 15, p ≤ 0.05). Histology showed necrosis and apoptosis (TUNEL assay) at the site of laser injury. At 24 h post-laser cardiac performance and VCt had recovered fully to control levels. Pretreatment with the cell-cycle inhibitor, aphidicolin, significantly inhibited functional recovery of the ventricle accompanied by a significant inhibition of cardiomyocyte proliferation. Conclusions: Laser-targeted injury of the zebrafish embryonic heart is a novel and reproducible model of cardiac injury and repair suitable for pharmacological and molecular studies.

Citation

Matrone, G., Taylor, J. M., Wilson, K. S., Baily, J., Love, G. D., Girkin, J. M., …Denvir, M. A. (2013). Laser-targeted ablation of the zebrafish embryonic ventricle: A novel model of cardiac injury and repair. International Journal of Cardiology, 168(4), 3913-3919. https://doi.org/10.1016/j.ijcard.2013.06.063

Journal Article Type Article
Publication Date Oct 9, 2013
Deposit Date May 22, 2014
Publicly Available Date Mar 28, 2024
Journal International Journal of Cardiology
Print ISSN 0167-5273
Publisher Elsevier
Peer Reviewed Peer Reviewed
Volume 168
Issue 4
Article Number 3913-3919
Pages 3913-3919
DOI https://doi.org/10.1016/j.ijcard.2013.06.063
Keywords Zebrafish, Heart, Laser; Injury, Repair.

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Copyright Statement
This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.





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