Arnaout Rima, Reischauer Sven, Stainier Didier Y R
Cardiovascular Research Institute and Division of Cardiology, University of California San Francisco;
Department for Biochemistry and Biophysics, University of California San Francisco; Max-Planck Institute for Heart and Lung Research.
J Vis Exp. 2014 Dec 12(94):52248. doi: 10.3791/52248.
Use of the zebrafish model system for studying development, regeneration, and disease is expanding toward use of adult hearts for cell dissociation and purification of RNA, DNA, and proteins. All of these applications demand the rapid recovery of significant numbers of zebrafish hearts to avoid gene regulatory, metabolic, and other changes that begin after death. Adult zebrafish hearts are also required for studying heart structure for a variety of mutants and for studying heart regeneration. However, the traditional zebrafish heart dissection is slow and difficult and requires specialized tools, making large-scale dissection of adult zebrafish hearts tedious. Traditional methods also harbor the risk of damaging the heart during the dissection. Here, we describe a method for dissection of adult zebrafish hearts that is fast, reproducible, and preserves heart architecture. Furthermore, this method does not require specialized tools, is painless for the zebrafish, can be performed on fresh or fixed specimens, and can be performed on zebrafish as young as one month old. The approach described expands the use of adult zebrafish for cardiovascular research.
斑马鱼模型系统在发育、再生和疾病研究中的应用正朝着利用成年斑马鱼心脏进行细胞解离以及RNA、DNA和蛋白质的纯化方向发展。所有这些应用都需要快速回收大量斑马鱼心脏，以避免死亡后开始出现的基因调控、代谢和其他变化。研究各种突变体的心脏结构以及心脏再生也需要成年斑马鱼心脏。然而，传统的斑马鱼心脏解剖过程缓慢且困难，需要专门的工具，这使得大规模解剖成年斑马鱼心脏变得繁琐。传统方法在解剖过程中还存在损伤心脏的风险。在此，我们描述一种成年斑马鱼心脏解剖方法，该方法快速、可重复且能保留心脏结构。此外，此方法不需要专门工具，对斑马鱼无痛，可在新鲜或固定标本上进行，并且可在仅一个月大的斑马鱼上操作。所描述的方法扩展了成年斑马鱼在心血管研究中的应用。
