PUBLICATION

A Possible Zebrafish Model of Polycystic Kidney Disease: Knockdown of wnt5a Causes Cysts in Zebrafish Kidneys

Authors
Huang, L., Xiao, A., Wecker, A., McBride, D.A., Choi, S.Y., Zhou, W., Lipschutz, J.H.
ID
ZDB-PUB-141210-11
Date
2014
Source
Journal of visualized experiments : JoVE   (94): (Journal)
Registered Authors
Xiao, An, Zhou, Weibin
Keywords
Medicine, Wnt5a, polycystic kidney disease, morpholino, microinjection, zebrafish, pronephros
MeSH Terms
  • Green Fluorescent Proteins/genetics
  • Polycystic Kidney Diseases/genetics*
  • Polycystic Kidney Diseases/metabolism
  • Oligonucleotides, Antisense/genetics
  • Disease Models, Animal*
  • Animals, Genetically Modified
  • Animals
  • Gene Knockdown Techniques
  • Wnt Proteins/deficiency
  • Wnt Proteins/genetics*
  • Wnt Proteins/metabolism
  • Zebrafish
(all 12)
PubMed
25489842 Full text @ J. Vis. Exp.
Abstract
Polycystic kidney disease (PKD) is one of the most common causes of end-stage kidney disease, a devastating disease for which there is no cure. The molecular mechanisms leading to cyst formation in PKD remain somewhat unclear, but many genes are thought to be involved. Wnt5a is a non-canonical glycoprotein that regulates a wide range of developmental processes. Wnt5a works through the planar cell polarity (PCP) pathway that regulates oriented cell division during renal tubular cell elongation. Defects of the PCP pathway have been found to cause kidney cyst formation. Our paper describes a method for developing a zebrafish cystic kidney disease model by knockdown of the wnt5a gene with wnt5a antisense morpholino (MO) oligonucleotides. Tg(wt1b:GFP) transgenic zebrafish were used to visualize kidney structure and kidney cysts following wnt5a knockdown. Two distinct antisense MOs (AUG - and splice-site) were used and both resulted in curly tail down phenotype and cyst formation after wnt5a knockdown. Injection of mouse Wnt5a mRNA, resistant to the MOs due to a difference in primary base pair structure, rescued the abnormal phenotype, demonstrating that the phenotype was not due to "off-target" effects of the morpholino. This work supports the validity of using a zebrafish model to study wnt5a function in the kidney.
Genes / Markers
Figures
No images available
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Expression
No data available
Phenotype
Fish Conditions Stage Phenotype Figure
li1Tg + MO2-wnt5acontrolLong-pec
li1Tg + MO2-wnt5acontrolProtruding-mouth
li1Tg + MO2-wnt5acontrolProtruding-mouth
li1Tg + MO2-wnt5acontrolProtruding-mouth
li1Tg + MO2-wnt5acontrolProtruding-mouth
li1Tg + MO2-wnt5acontrolProtruding-mouth
li1Tg + MO2-wnt5acontrolProtruding-mouth
li1Tg + MO2-wnt5acontrolProtruding-mouth
li1Tg + MO2-wnt5acontrolProtruding-mouth
li1Tg + MO3-wnt5acontrolLong-pec
1 - 10 of 15
Show
Mutations / Transgenics
Allele Construct Type Affected Genomic Region
li1TgTransgenic Insertion
    1 - 1 of 1
    Show
    Human Disease / Model
    Human Disease Fish Conditions Evidence
    polycystic kidney diseaseTAS
    1 - 1 of 1
    Show
    Sequence Targeting Reagents
    Target Reagent Reagent Type
    wnt5aMO2-wnt5aMRPHLNO
    wnt5aMO3-wnt5aMRPHLNO
    1 - 2 of 2
    Show
    Fish
    Antibodies
    Orthology
    Engineered Foreign Genes
    Marker Marker Type Name
    EGFPEFGEGFP
    1 - 1 of 1
    Show
    Mapping
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    Citation
    Huang, L., Xiao, A., Wecker, A., McBride, D.A., Choi, S.Y., Zhou, W., Lipschutz, J.H. (2014) A Possible Zebrafish Model of Polycystic Kidney Disease: Knockdown of wnt5a Causes Cysts in Zebrafish Kidneys. Journal of visualized experiments : JoVE. (94).