ZFIN ID: ZDB-PUB-160904-3
Monitoring Wnt Signaling in Zebrafish Using Fluorescent Biosensors
Facchinello, N., Schiavone, M., Vettori, A., Argenton, F., Tiso, N.
Date: 2016
Source: Methods in molecular biology (Clifton, N.J.)   1481: 81-94 (Chapter)
Registered Authors: Argenton, Francesco, Facchinello, Nicola, Schiavone, Marco, Tiso, Natascia, Vettori, Andrea
Keywords: Chemical biology, Transgenic reporter, Transposase, Zebrafish
MeSH Terms:
  • Animals
  • Animals, Genetically Modified
  • Biosensing Techniques/methods*
  • Enhancer Elements, Genetic*
  • Gene Expression Regulation, Developmental
  • Genes, Reporter
  • Green Fluorescent Proteins/genetics
  • Microinjections/methods
  • Promoter Regions, Genetic*
  • Wnt Proteins/genetics*
  • Wnt Proteins/isolation & purification
  • Wnt Signaling Pathway/genetics
  • Zebrafish/genetics
PubMed: 27590154 Full text @ Meth. Mol. Biol.
In this chapter, we are presenting methods to monitor and quantify in vivo canonical Wnt signaling activities at single-cell resolution in zebrafish. Our technology is based on artificial enhancers, obtained by polymerization of TCF binding elements, cloned upstream to ubiquitous or tissue-specific promoters. The different promoter/enhancer combinations are used to drive fluorescent protein reporter constructs integrated in the zebrafish germline by microinjection of fertilized zebrafish eggs. Fish with a single integration site are selected by Mendelian analysis of fluorescent carriers, and heterozygous offspring are used to monitor and quantify canonical Wnt activities. Open source public domain software such as ImageJ/Fiji is used to calculate the integrated densities in the region of interest and compare the effect of experimental conditions on control and treated animals.