Knockdown of rasip1 in zebrafish affects vascular integrity. (A-B) Tg(kdrl:EGFP)^s843 zebrafish embryos at 26 hpf were injected with control (ctrl) morpholino (MO) (A) or rasip1 MO (B). Arrows indicate dorsal aorta. Arrowheads indicate PCV. Insets show magnification of trunks. Note stalling of intersomitic vessels (ISVs). (C-F) Confocal projections of trunk axial vessels from control (C,E) or rasip1 (D,F) MO-injected Tg(kdrl:EGFP)^s843;Tg(gata1:DsRED)^sd2 double transgenic live embryos at 48 hpf. Images are maximum projections of 3 confocal slices. Note irregular appearance of the rasip1 KD DA and PCV (green) (D,F), which have patent lumen and contain circulating RBCs (red). (G-H) Maximum projections of head vasculature in ctrl (G) and rasip1 (H) MO-injected double transgenic fish at 48 hpf. Intralumenal RBCs were observed in both embryos (insets), whereas extravascular RBCs were only seen in the rasip1 KD embryo. The streaking appearance of red signal in panels C-F is due to rapid movement of RBCs. Scale bars: (A-B) 100 μm; (C-H) 20 μm.