386A Poster - 04. Stem cells, regeneration and tissue injury
Thursday April 07, 2:00 PM - 4:00 PM

Authors:
Gary Teeters; Sarah Siegrist

Affiliation: University of Virginia

Keywords:
k. regeneration; q. other (Cell Ablation)

Targeted cell ablation is a common method used to assay cell function during development and to determine cellular limits during regeneration. Many techniques are available to achieve cell ablation, yet improvements are still needed to increase spatial and temporal control. One method commonly used in the Zebrafish, Danio rerio is a Nitroreductase (NTR) mediated ablation system. In this technique cells expressing NTR can be induced to undergo cell death following drug treatment. For example, when metronidazole (MTZ) is exposed to cells expressing Nitroreductase, NTR converts MTZ into a DNA cross linker inducing cells to die. We set out to determine whether this commonly used cell ablation system could be used in Drosophila. We cloned the Nitroreductase gene from Escherichia Coli used in Curado et al. 2007 with a GFP reporter to examine cell death in vivo. This technique would allow for greater temporal control of cell ablation without the need of temperature sensitive Gal80 or mutant alleles instead by regulating exposure to nitroimidazole antibiotics. This project means to adapt the protocols from Danio rerio to Drosophila melanogaster which will modify the exposure method, concentration, and time. We aim to show that exposure to nitroimidazole antibiotics through ingestion at sub toxic levels will be able to ablate cells in Gal4 specific manner with temporal control and no off-target toxicity.