title: Validation of nitroreductase, a prodrug-activating enzyme, mediated cell death in embryonic zebrafish (Danio rerio).
abstract: 1Escherichia coli gene nfsB encodes a nitroreductase (NTR) enzyme that converts prodrugs like metronidazole (Met) and CB 1954 to cytotoxic metabolites. My coworkers and I have validated this prodrug-enzyme system in zebrafish (Danio rerio) by ubiquitously expressing NTR-EGFP fusion protein and exposing these embyos to CB 1954 and Met. These embryos showed extensive gross pathologic changes and death by 24 h of incubation in the prodrugs. They also exhibited widespread and marked apoptotic changes by 8 h of incubation in Met. Neither the prodrugs themselves nor the NTR-EGFP fusion protein were toxic to the developing zebrafish embryos. Thus the NTR-CB 1954 and NTR-Met systems can be used to ablate a wide variety of cells and tissues in zebrafish embryos.
authors.affiliation: Department of Molecular and Comparative Pathobiology, Johns Hopkins University, Baltimore, MD, USA. hpishar1@jhmi.edu
authors.forename: Harshan
authors.initials: H
authors.lastname: Pisharath
mesh_terms: D000818:Animals; D001388:Aziridines / Q000494:pharmacology; D016923:Cell Death; D003001:Cloning, Molecular; D004625:Embryo, Nonmammalian / Q000201:enzymology / Q000473:pathology*; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects*; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics*; D020287:In Situ Nick-End Labeling; D008795:Metronidazole / Q000494:pharmacology; D018613:Microscopy, Confocal; D009601:Nitroreductases / Q000235:genetics*; D030781:Organisms, Genetically Modified; D016133:Polymerase Chain Reaction; D011355:Prodrugs / Q000494:pharmacology*; D011993:Recombinant Fusion Proteins / Q000235:genetics; D015027:Zebrafish / Q000196:embryology*
chemical_list: D001388:Aziridines; D011355:Prodrugs; D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; D008795:Metronidazole; D049452:Green Fluorescent Proteins; C100099:tretazicar; D009601:Nitroreductases