An ATF2-based luciferase reporter to monitor non-canonical Wnt signaling in Xenopus embryos.
Non-canonical/planar cell polarity (PCP) Wnt signaling plays important roles in embryonic development and tissue homeostasis, and is implicated in human disease. Monitoring Wnt/PCP signaling relies mostly on semi-quantitative bioassays or biochemical analysis. Here we describe a luciferase reporter assay based on an ATF2 response element, which faithfully monitors non-canonical Wnt signaling in Xenopus embryos. The assay is simple, quantitative, and robust. It can be used to detect non-canonical Wnt signaling changes following gain and loss of function of pathway components, including Wnt, Frizzled, Ror2, Disheveled, Rac1, MKK7, and JNK. Wnt/PCP signaling has recently been implicated in left-right asymmetry and our reporter assay suggests that in gastrula embryos there is a right-ward bias in Wnt/PCP signaling. We also mapped Wnt/PCP signaling in the early Xenopus embryo and find that it peaks in the dorso-vegetal region, paralleling Wnt/β-catenin signaling.
Division of Molecular Embryology, DKFZ-ZMBH Alliance, Deutsches Krebsforschungszentrum, Heidelberg, Germany.
Bisei
B
Ohkawara
Christof
C
Niehrs
D051698:Activating Transcription Factor 2 / Q000235:genetics*; D000818:Animals; D030801:Animals, Genetically Modified; D058948:Cell Tracking / Q000379:methods; D004625:Embryo, Nonmammalian; D047108:Embryonic Development / Q000235:genetics / Q000502:physiology; D018507:Gene Expression Regulation, Developmental; D017930:Genes, Reporter / Q000502:physiology; D008156:Luciferases / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics; D015398:Signal Transduction / Q000235:genetics; D051153:Wnt Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D014981:Xenopus / Q000196:embryology* / Q000235:genetics* / Q000378:metabolism
D051698:Activating Transcription Factor 2; D011993:Recombinant Fusion Proteins; D051153:Wnt Proteins; D008156:Luciferases