wwtr1-MO injection effect on endothelial marker expression and vascular tree morphology. WISH analysis with riboprobes against fli-1 and tie-1 mRNA at 48 hpf (A–D), and qPCR analysis where fold change is relative to rpl13a (E and F), show comparable results in uninjected (A and C) and standard control-MO (scMO) zebrafish (48 hpf). qPCR results are the average of two independent experiments conducted on pools of 50 embryos each (p > 0.01). Zebrafish injected with standard control-MO (dark gray) and wwtr1-MO (light gray), H, I). Scale bar = 200 µm. Anterior is to the left in lateral view. (G-I) Vasculature of wwtr1 morphant larvae is altered in the pharyngeal region of tg(kdr-like:gfp) zebrafish (4 dpf), with patterning defects and loss of blood vessels. Pharyngeal vessels and thyroid follicles were immunolabeled for GFP (green) and T4 antibody (red) in 4 dpf larvae. Uninjected (G) and wwtr1-MO injected (H and I). Arrows indicate bilateral ventral aorta (H and I). (Branchial) aortic arch vessels are either reduced and/or dysplastic in wwtr1 morphants. Arrowhead marks thyroid follicle on a (branchial) aortic arch artery of morphant larvae (I). Scale bar = 200 µm. AA, (branchial) aortic arch artery; AA1, first (branchial) aortic arch artery; AA2, second (branchial) aortic arch artery; AA3, third (branchial) aortic arch artery; AA4, fourth (branchial) aortic arch artery; HA, hypobranchial arch artery; OFT, outflow tract; ORA, opercular artery; VA, ventral aorta. Anterior is to the top in ventral view. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)
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