Involvement of AP-2rep in morphogenesis of the axial mesoderm in Xenopus embryo

We have previously isolated a cDNA clone coding for Xenopus AP-2rep (activator protein-2 repressor), a member of the Krüppel-like factor family, and reported its expression pattern in developing Xenopus embryos. In the present study, the physiological function of AP-2rep in the morphogenetic movements of the dorsal mesoderm and ectoderm was investigated. Embryos injected with either AP-2rep or VP16repC (a dominant-negative mutant) into the dorsal marginal zone at the 4-cell stage exhibited abnormal morphology in dorsal structures. Both AP-2rep and VP16repC also inhibited the elongation of animal cap explants treated with activin without affecting the expression of differentiation markers. Whole-mount in situ hybridization analysis revealed that expression of brachyury and Wnt11 was greatly suppressed by injection of VP16repC or AP-2rep morpholino, but expression was restored by the simultaneous injection of wild-type AP-2rep RNA. Furthermore, the morphogenetic abnormality induced by injection of VP16repC or AP-2rep morpholino was restored by simultaneous injection of brachyury or Wnt11 mRNA. These results show that AP-2rep is involved in the morphogenesis of the mesoderm at the gastrula stage, via the brachyury and/or Wnt pathways.