Vascular analysis in Tg(fli1:egfp)^y1 perlecan morphant embryos. (A and B) Epifluorescence microscopy with 3D deconvolution comparing the trunk vasculature of 2-dpf control versus MO-DI embryos. Note the correct formation of DLAV, ISV, DA, and PCV in the control (A). In contrast, the perlecan morphant (B) exhibits abnormal ISVs. Often the ISV sprouts fail to completely migrate along the myoseptae and to anastomose (arrows). (C and D) Merged images corresponding to those shown in A and B taken with both fluorescence deconvolution and DIC microscopy. Note that the ISVs barely reach the notochord (NC) and do not properly migrate to the dorsal region, thereby failing to interconnect and properly form the DLAV. (E and F) Lateral and dorsal views of alkaline phosphatase–stained control embryos at 3 dpf. Notice the well-developed SIV. (G–I) Three representative MO-DI perlecan morphants showing the complete absence of SIV and reduced alkaline phosphatase staining throughout the trunk vasculature and head region. Notice that J is a higher magnification of G. The only detectable signal is present along the axial vessels (arrows in G and J). Bars, 500 μm.