Real-time observation of Wnt beta-catenin signaling in the chick embryo.
A critical mediator of cell-cell signaling events during embryogenesis is the highly conserved Wnt family of secreted proteins. Reporter constructs containing multimerized TCF DNA binding sites have been used to detect Wnt beta-catenin dependent activity during animal development. In this report, we have constructed and compared several TCF green fluorescent protein (GFP) reporter constructs. They contained 3, 8, or 12 TCF binding sites upstream of a minimal promoter driving native or destabilized enhanced GFP (EGFP). We have used the electroporation of somites in the chick embryo as a paradigm to test them in vivo. We have verified that they all respond to Wnt signaling in vivo. We have then assessed their efficiency at reflecting the activity of the Wnt pathway. Using destabilized EGFP reporter constructs, we show that somite cells dynamically regulate Wnt/beta-catenin-dependent signaling, a finding that was confirmed by performing time-lapse video confocal observation of electroporated embryos.
Rios
Anne C
AC
Developmental Biology Institute of Marseille Luminy (IBDML), CNRS UMR 6216, Université de la Méditerranée, Campus de Luminy, Marseille, France.
Denans
Nicolas
N
Marcelle
Christophe
C
D000818:Animals; D001665:Binding Sites / Q000235:genetics; D002450:Cell Communication / Q000502:physiology*; D002642:Chick Embryo; D018274:Electroporation; D049452:Green Fluorescent Proteins / Q000378:metabolism; D018613:Microscopy, Confocal; D015398:Signal Transduction / Q000502:physiology*; D019170:Somites / Q000166:cytology / Q000378:metabolism*; D051820:TCF Transcription Factors / Q000378:metabolism*; D014741:Video Recording; D051153:Wnt Proteins / Q000378:metabolism*; D051176:beta Catenin / Q000378:metabolism*
D051820:TCF Transcription Factors; D051153:Wnt Proteins; D051176:beta Catenin; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins