Laser-targeted ablation of the zebrafish embryonic ventricle: a novel model of cardiac injury and repair.
Laser-targeted ablation of the zebrafish embryonic ventricle: a novel model of cardiac injury and repair.
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DOI：
10.1016/j.ijcard.2013.06.063
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发表时间：
2013-10-09
影响因子：
3.5
通讯作者：
Denvir, Martin A.
中科院分区：
文献类型：
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作者：
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.
虽然成年斑马鱼(Danio Rerio)的心脏在心尖切除后表现出非凡的自我更新能力，但对胚胎心脏对损伤的反应知之甚少。利用表达心肌细胞特异性绿色荧光蛋白的转基因斑马鱼，用激光诱导跳动中的斑马鱼胚胎心脏损伤。观察激光照射后2、24、48h的射血分数(EF)、心率(HR)和尾静脉血流量(CVBF)的变化。用激光共聚焦显微镜分别计数DAPI(VCT)和磷酸组蛋白H3(VCM)阳性核，观察激光损伤后心肌细胞总数和有丝分裂数的变化。激光对脑室的损伤导致心动过缓和轻微的心包出血。在激光损伤后2小时，激光心脏的心功能较对照组显著降低(HR 117±0.11 vs 167±9.9μ/min，p<≤=0.001；EF 14.1±1.8 vs 20.1±1.3%，p>≤=0.001；CVBF 103±±0.15 vs 316±0.13μms−：1，p>≤(0.001))。离体心在激光照射后2小时的VCT与对照组相比显著降低(195.00±0.15vs238±0.15，p>≤<0.05)。组织学显示激光损伤部位有坏死和细胞凋亡(TUNEL法)。在激光治疗后24小时，心功能和VCT已完全恢复到对照水平。细胞周期抑制剂aphidiclin可显著抑制心室功能的恢复，同时明显抑制心肌细胞的增殖。激光靶向损伤斑马鱼胚胎心脏是一种新的、可重复性的心脏损伤和修复模型，适用于药理学和分子生物学研究。
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. 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. 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. 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.
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