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冠状动脉功能障碍诱导的心脏损伤后斑马鱼心脏的再生

Zebrafish heart regeneration after coronary dysfunction-induced cardiac damage.

作者信息

Sun Jisheng, Peterson Elizabeth A, Jiao Cheng, Chen Xin, Zhao Yun, Wang Jinhu

机构信息

Division of Cardiology, School of Medicine, Emory University, Atlanta, GA, 30322, USA.

Division of Cardiology, School of Medicine, Emory University, Atlanta, GA, 30322, USA.

出版信息

Dev Biol. 2022 Jul;487:57-66. doi: 10.1016/j.ydbio.2022.04.008. Epub 2022 Apr 29.

Abstract

Over the past 20 years, various zebrafish injury models demonstrated efficient heart regeneration after cardiac tissue loss. However, no established coronary vessel injury methods exist in the zebrafish model, despite coronary endothelial dysfunction occurring in most patients with acute coronary syndrome. This is due to difficulties performing surgery on small coronary vessels and a lack of genetic tools to precisely manipulate coronary cells in zebrafish. We determined that the Notch ligand gene deltaC regulatory sequences drive gene expression in zebrafish coronary endothelial cells, enabling us to overcome these obstacles. We created a deltaC fluorescent reporter line and visualized robust coronary growth during heart development and regeneration. Importantly, this reporter facilitated the visualization of coronary growth without an endocardial background. Moreover, we visualized robust coronary growth on the surface of juvenile hearts and regrowth in the wounded area of adult hearts ex vivo. With this approach, we observed growth inhibition by reported vascular growth antagonists of the VEGF, EGF and Notch signaling pathways. Furthermore, we established a coronary genetic ablation system and observed that severe coronary endothelial cell loss resulted in fish death, whereas fish survived mild coronary cell loss. Coronary cell depletion triggered regenerative responses, which resulted in the restoration of damaged cardiac tissues within several weeks. Overall, our work demonstrated the efficacy of using deltaC regulatory elements for high-resolution visualization of the coronary endothelium; screening small molecules for coronary growth effects; and revealed complete recovery in adult zebrafish after coronary-induced heart damage.

摘要

在过去20年里,各种斑马鱼损伤模型都证明了心脏组织受损后心脏具有高效的再生能力。然而,斑马鱼模型中尚无成熟的冠状动脉损伤方法,尽管大多数急性冠状动脉综合征患者都存在冠状动脉内皮功能障碍。这是由于在小型冠状动脉上进行手术存在困难,且缺乏在斑马鱼中精确操纵冠状动脉细胞的基因工具。我们确定Notch配体基因deltaC的调控序列可驱动斑马鱼冠状动脉内皮细胞中的基因表达,从而使我们能够克服这些障碍。我们创建了一个deltaC荧光报告系,并观察到心脏发育和再生过程中冠状动脉的强劲生长。重要的是,该报告系有助于在没有心内膜背景的情况下观察冠状动脉的生长。此外,我们在幼鱼心脏表面观察到了强劲的冠状动脉生长,并在成鱼心脏伤口区域的离体心脏中观察到了冠状动脉的再生。通过这种方法,我们观察到了VEGF、EGF和Notch信号通路中已报道的血管生长拮抗剂对生长的抑制作用。此外,我们建立了一种冠状动脉基因消融系统,观察到严重的冠状动脉内皮细胞损失会导致鱼类死亡,而轻度冠状动脉细胞损失的鱼类能够存活。冠状动脉细胞耗竭引发了再生反应,导致受损心脏组织在数周内得以恢复。总体而言,我们的工作证明了使用deltaC调控元件对冠状动脉内皮进行高分辨率可视化、筛选影响冠状动脉生长的小分子的有效性,并揭示了成年斑马鱼在冠状动脉损伤后心脏的完全恢复情况。

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