Cav-1 regulates zebrafish dorsoventral patterning via maternal Wnt signaling. (A) Expression patterns of reporter genes (indicated on the top) for FGF, Nodal, BMP and Wnt signalings in wild-type (WT) or embryos injected with 300 pg cav-1α mRNA, 300 pg cav-1β mRNA, 10 ng cav-1α-MO or 10 ng cav-1β-MO. Developmental stages are shown at the bottom. Lateral views of embryos at 12 hpf with anterior toward the top for sef. Dorsal views of embryos at shield stage with animal pole toward the top for spry4 and boz. Animal pole views of embryos at shield stage with the dorsal toward the right for xbp1, flh and dkk1. Animal pole views of embryos at 30% epiboly stage with the dorsal toward the right for squint (sqt). Animal pole views of embryos at 70% epiboly stage with the dorsal toward the right for ved and vent. Arrows point to the margins of expressed ved and vent. Dorsoanterior views of embryos at 12 hpf with the anterior toward the top for otx2. Vegetal views of embryos at 80% epiboly stage with dorsal toward the right for wnt8a. Dorsal views of embryos at 12 hpf with the anterior toward the top for tbx6. (B) Total number of embryos examined (indicated at the bottom) and percentage of embryos with decreased expression of genes sqt, dkk1, boz and otx2 in (A) after injection of cav-1 mRNA. (C) Total number of embryos examined (indicated at the bottom) and percentage of embryos with increased expression of genes sqt, dkk1, boz and otx2 in (A) after injection of cav-1-MOs. (D) Luciferase activity of Top-flash reporter for Wnt signaling was inhibited by injection of 300 pg cav-1α or -1β mRNA, but enhanced by injection of 10 ng cav-1α-MO or 10 ng cav-1β-MO. Injection of 50 pg mRNA for constitutively active β-catenin-1 (β-cat mRNA) stimulated activity of Top-flash reporter and was used as the positive control. Data for luciferase assays represent mean ± S.D. from three independent experiments. *indicates p < 0.05 and **indicates p < 0.01 vs. Top-flash-injected embryos.
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