title: A zebrafish model of conditional targeted podocyte ablation and regeneration.
abstract: Podocytes are specialized cells that contribute critically to the normal structure and function of the glomerular filtration barrier. Their depletion plays an important role in the pathogenesis of glomerulosclerosis. Here, we report generation of a genetic model of conditional podocyte ablation and regeneration in zebrafish using a bacterial nitroreductase strategy to convert a prodrug, metronidazole, into a cytotoxic metabolite. A transgenic zebrafish line was generated that expresses green fluorescence protein (GFP) and the nitroreductase fusion protein under the control of the podocin promoter Tg(podocin:nitroreductase-GFP). Treatment of these transgenic zebrafish with metronidazole results in podocyte apoptosis, a loss of nephrin and podocin expression, foot process effacement, and a leaky glomerular filtration barrier. Following metronidazole washout, proliferating cells were detected in the glomeruli of recovering transgenic fish with a restoration of nitroreductase-GFP fluorescence, nephrin and podocin expression, a reestablishment of normal foot process architecture, and glomerular barrier function. Thus, our studies show that zebrafish podocytes are capable of regenerating following depletion, and establish the Tg(podocin:NTR-GFP) fish as a new model to study podocyte injury and repair.
authors: Center for Systems Biology, Program in Membrane Biology, Division of Nephrology, Department of Medicine, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts 02114, USA.
authors: Jianmin
authors: J
authors: Huang
authors: Mary
authors: M
authors: McKee
authors: Hong D
authors: HD
authors: Huang
authors: Alice
authors: A
authors: Xiang
authors: Alan J
authors: AJ
authors: Davidson
authors: Hua A J
authors: HA
authors: Lu
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D017209:Apoptosis / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D005919:Glomerular Filtration Rate / Q000187:drug effects; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D008795:Metronidazole / Q000378:metabolism / Q000633:toxicity; D051379:Mice; D023421:Models, Animal; D009601:Nitroreductases / Q000235:genetics / Q000378:metabolism; D010539:Permeability; D050199:Podocytes / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D011355:Prodrugs / Q000378:metabolism / Q000633:toxicity; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012038:Regeneration / Q000187:drug effects; D015027:Zebrafish / Q000235:genetics / Q000378:metabolism; D029961:Zebrafish Proteins / Q000378:metabolism
chemical_list: D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; C407223:NPHS2 protein; D011355:Prodrugs; D011993:Recombinant Fusion Proteins; D029961:Zebrafish Proteins; C113267:nephrin; D008795:Metronidazole; D049452:Green Fluorescent Proteins; D009601:Nitroreductases