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

Matrone, Gianfranco and Taylor, Jonathan M. and Wilson, Kathryn S. and Baily, James and Love, Gordon D. and Girkin, John M. and Mullins, John J. and Tucker, Carl S. and Denvir, Martin A. (2013) 'Laser-targeted ablation of the zebrafish embryonic ventricle : a novel model of cardiac injury and repair.', International journal of cardiology., 168 (4). pp. 3913-3919.

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.

Item Type:Article
Keywords:Zebrafish, Heart, Laser; Injury, Repair.
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Status:Peer-reviewed
Publisher Web site:http://dx.doi.org/10.1016/j.ijcard.2013.06.063
Publisher 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.
Date accepted:No date available
Date deposited:30 May 2014
Date of first online publication:October 2013
Date first made open access:No date available

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