Fig. 4

Morpholino antisense knockdown of zNXT2 in transgenic zebrafish embryos expressing cmlc2-GFP and flk-RFP. zNXT2 knockdown causes pericardial edema (compare A, C, E, G with B, D, F, H), abnormal relative positions of two chambers and chamber dilation (compare insets in C-H showing CMLC2 promoter driven GFP expression patterns in myocardium of atriums and ventricles). Observation of endothelial cells at the atrioventricular boundary in living flk-1- RFP transgenic embryos allowing visualization of cardiac valve formation (I and J). Clustering of endothelial cells was clearly visible in the cardiac valve region in wild-type embryos (I, arrow) whereas injection of zNXT2 morpholino caused a failure of clustering of endocardial cells at the atrioventricular boundary (J). All embryos are lateral view, anterior to the left. a: atrium; v: ventricle; OT: outflow tract; A, C, E, G: uninjected wild-type embryos; B, D, F, H: morpholino-injected embryos. wt: wild type; dpf: days post fertilization; MO: zNXT2 morpholino.