Conditional targeted cell ablation in zebrafish: A new tool for regeneration studies
Conditional targeted cell ablation in zebrafish: A new tool for regeneration studies
复制标题
DOI：
10.1002/dvdy.21100
复制
发表时间：
2007-04-01
影响因子：
2.5
通讯作者：
Stainier, Didier Y. R.
中科院分区：
文献类型：
--
作者：
Curado, Silvia;Anderson, Ryan M.;Stainier, Didier Y. R.
斑马鱼中的条件性靶向细胞消融将极大地扩展该遗传模型系统在发育和再生研究中的实用性，因为其具有广泛的再生能力。在这里，我们表明，通过结合化学和遗传工具，可以消融细胞的时间和空间特异性的方式在斑马鱼幼虫。为此，我们使用细菌硝基还原酶（NTR）酶转化前药甲硝唑（Mtz）成为一种细胞毒性的DNA交联剂。为了研究该系统的效率，我们针对心脏、胰腺和肝脏中的三种不同细胞系。在组织特异性启动子的控制下表达融合蛋白青色荧光蛋白-NTR（CFP-NTR）允许我们在特定时间诱导心肌细胞、胰腺β细胞和肝细胞的死亡。此外，我们已经观察到Mtz可以被有效地洗掉，并且在Mtz撤出后，严重受影响的组织可以迅速恢复。这些研究结果表明，NTR/Mtz系统是有效的时间和空间控制的细胞消融在斑马鱼，从而构成一个最有前途的遗传工具，分析组织的相互作用以及再生的机制。
Conditional targeted cell ablation in zebrafish would greatly expand the utility of this genetic model system in developmental and regeneration studies, given its extensive regenerative capabilities. Here, we show that, by combining chemical and genetic tools, one can ablate cells in a temporal- and spatial-specific manner in zebrafish larvae. For this purpose, we used the bacterial Nitroreductase (NTR) enzyme to convert the prodrug Metronidazole (Mtz) into a cytotoxic DNA cross-linking agent. To investigate the efficiency of this system, we targeted three different cell lineages in the heart, pancreas, and liver. Expression of the fusion protein Cyan Fluorescent Protein-NTR (CFP-NTR) under control of tissue-specific promoters allowed us to induce the death of cardiomyocytes, pancreatic beta-cells, and hepatocytes at specific times. Moreover, we have observed that Mtz can be efficiently washed away and that, upon Mtz withdrawal, the profoundly affected tissue can quickly recover. These findings show that the NTR/Mtz system is effective for temporally and spatially controlled cell ablation in zebrafish, thereby constituting a most promising genetic tool to analyze tissue interactions as well as the mechanisms underlying regeneration.