Prodrug-Induced Cell Ablation in Zebrafish

Transgenic expression of the bacterial enzyme nitroreductase (nfsB, NTR) facilitates prodrug-inducible cell-type specific ablation in zebrafish [29] (link), [35] (link). Transgenic lines expressing NTR and a fluorescent reporter in specific cell types were used to determine whether ARQiv could detect the selective loss and regeneration of fluorescently labeled cells. Prior to cell ablation, NTR-expressing and control non-expressing larvae were scanned to determine pre-treatment fluorescence and background levels. Afterwards, larvae were treated for 24 hrs with media containing the prodrug metronidazole (MTZ, 10 mM for pancreatic beta cell ablation, 2.5 mM for rod photoreceptor cell ablation) to induce ablation of NTR-expressing cells, or with control media. After 24 hrs, all wells were rinsed three times and returned to control media. Subsequent post-treatment and recovery scans were performed at the indicated intervals. Confocal imaging was used to verify the loss and return of fluorescent reporters in the targeted cell types in identically treated subjects.