Fig. 1

Morpholino knockdown of ccm3a/b in zebrafish embryos causes cardiovascular defects distinct from ccm1/2 mutants. (A) Light images of ccm3a/b morpholino injected embryos at 54 hpf. Morphant embryos display a severe pericardial edema and complete circulatory block, but no gross morphological defects when injected with 0.25–0.75 ng ccm3a/b ATG morpholino. (B) Fluorescent microscopy images of ccm3a/b morpholino injected and non-injected double transgenic kdrl:EGFP; gata1:dsRED embryos at 54 hpf. Morphants lack circulation with blood pooling at the yolk. (C) Confocal images of the ccm3a/b morpholino injected Tg(myl7:EGFP)^twu34 wildtype and ccm2 (valentine) mutant embryos. At 54 hpf, ccm3a/b morphant hearts are dysmorphic but not enlarged, unlike the ccm2 (valentine) mutants whose hearts appear grossly dilated. (D) Confocal images of ccm3a/b morpholino injected and non-injected double transgenic kdrl:EGFP, myl7:ras-cherry embryos. Injection of morpholino causes thinning of the ventral aorta and the first aortic arches by 54 hpf.