title: Conditional targeted cell ablation in zebrafish: a new tool for regeneration studies.
abstract: 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.
authors.affiliation: Department of Biochemistry and Biophysics, Cardiovascular Research Institute, University of California at San Francisco, San Francisco, California 94158-2324, USA.
authors.forename: Silvia
authors.initials: S
authors.lastname: Curado
authors.forename: Ryan M
authors.initials: RM
authors.lastname: Anderson
authors.forename: Benno
authors.initials: B
authors.lastname: Jungblut
authors.forename: Jeff
authors.initials: J
authors.lastname: Mumm
authors.forename: Eric
authors.initials: E
authors.lastname: Schroeter
authors.forename: Didier Y R
authors.initials: DY
authors.lastname: Stainier
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D004625:Embryo, Nonmammalian; D018507:Gene Expression Regulation, Developmental; D041722:Genes, Transgenic, Suicide; D005818:Genetic Engineering / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006321:Heart / Q000502:physiology; D022781:Hepatocytes / Q000502:physiology; D050417:Insulin-Secreting Cells / Q000502:physiology; D008954:Models, Biological; D009206:Myocardium / Q000166:cytology; D032383:Myocytes, Cardiac / Q000166:cytology; D009601:Nitroreductases / Q000235:genetics / Q000378:metabolism; D009928:Organ Specificity / Q000235:genetics; D010641:Phenotype; D011401:Promoter Regions, Genetic; D012038:Regeneration / Q000502:physiology*; D015027:Zebrafish / Q000196:embryology / Q000235:genetics / Q000502:physiology*
chemical_list: C485182:Cyan Fluorescent Protein; D049452:Green Fluorescent Proteins; D009601:Nitroreductases