PUBLICATION

Inducible Podocyte Injury and Proteinuria in Transgenic Zebrafish

Authors
Zhou, W., and Hildebrandt, F.
ID
ZDB-PUB-120327-5
Date
2012
Source
Journal of the American Society of Nephrology : JASN   23(6): 1039-1047 (Journal)
Registered Authors
Zhou, Weibin
Keywords
none
MeSH Terms
  • Animals
  • Animals, Genetically Modified*
  • Disease Models, Animal
  • Glomerular Filtration Barrier
  • Immunohistochemistry
  • Intracellular Signaling Peptides and Proteins/metabolism
  • Kidney Glomerulus/metabolism
  • Membrane Proteins/metabolism
  • Metronidazole/pharmacology
  • Microscopy, Confocal
  • Nephrotic Syndrome/metabolism
  • Nephrotic Syndrome/pathology*
  • Podocytes/drug effects*
  • Podocytes/metabolism
  • Proteinuria/physiopathology*
  • Sensitivity and Specificity
  • Vitamin D-Binding Protein/metabolism
  • Zebrafish/genetics*
PubMed
22440901 Full text @ J. Am. Soc. Nephrol.
Abstract

Damage or loss of podocytes causes glomerulosclerosis in murine models, and mutations in podocyte-specific genes cause nephrotic syndrome in humans. Zebrafish provide a valuable model for kidney research, but disruption of pronephroi leads to death within a few days, thereby preventing the study of CKD. In this study, we generated an inducible model of podocyte injury in zebrafish (pod::NTR-mCherry) by expressing a bacterial nitroreductase, which converts metronidazole to a cytotoxin, specifically in podocytes under the control of the zebrafish nphs2/podocin promoter. Application of the prodrug metronidazole to the transgenic fish induces acute damage to the podocytes in pronephroi of larval zebrafish and the mesonephroi of adult zebrafish, resulting in foot-process effacement and podocyte loss. We also developed a functional assay of the glomerular filtration barrier by creating transgenic zebrafish expressing green fluorescent protein (GFP)–tagged vitamin D–binding protein (VDBP) as a tracer for proteinuria. In the VDBP-GFP and pod::NTR-mCherry double-transgenic fish, induction of podocyte damage led to whole-body edema, and the proximal tubules reabsorbed and accumulated VDBP-GFP that leaked through the glomeruli, mimicking the phenotype of human nephrotic syndrome. Moreover, expression of wt1b::GFP, a marker for the developing nephron, extended into the Bowman capsule in response to podocyte injury, suggesting that zebrafish have a podocyte-specific repair process known to occur in mammalian metanephros. These data support the use of these transgenic zebrafish as a model system for studies of glomerular pathogenesis and podocyte regeneration.

Genes / Markers
Figures
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Expression
Phenotype
Mutation and Transgenics
Human Disease / Model Data
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping
Errata and Notes