Fig. 2

Essential regulation of CNS angiogenesis by Reck.

(A) Lateral views of skin pigmentation patterns of wild-type and gpr124^s984/s984 adults. Bottom panels are high magnification images of the upper panel boxed areas. (B) Dorsal views of DRGs in the trunk region of wild-type and gpr124^s984/s984 Tg(ngn1:GFP); Tg(sox10:mRPF) larvae at 72 hpf; arrows point to DRGs (anterior to the left). Scale bar, 50 µm. (C) Quantification of ngn1:GFP⁺ DRGs in 72 hpf wild-type, gpr124^s984/+ and gpr124^s984/s984 larvae. ngn1:GFP⁺ DRGs were counted on one side of the larvae. (D) Lateral views of DRG in Tg(ngn1:GFP);Tg(sox10:mRPF) wild-type and gpr124^s984/s984 animals. Scale bar, 20 µm. (E) Lateral views (anterior to the left) of Tg(kdrl:GFP) control and morphant embryos at 36 hpf (hindbrain region, upper panels) and 24 hpf (trunk region, bottom panels). Arrows point to the forming CtAs (upper panels) and ISVs (lower panels). Scale bar, 50 µm. (F) Maximal intensity projection of a confocal z-stack of Tg(kdrl:GFP) wild-type and reck morphant cranial vasculature at 60 hpf in dorsal views (anterior to the left; scale bar, 100 µm) and quantification of hindbrain CtAs after Reck downregulation by anti-sense morpholino injections at various doses. (G) Quantification of hindbrain CtAs in control and gpr124 or reck morphants at 60 hpf after injection at the one-cell stage of 100 pg RNA encoding Gpr124 or Reck. The insert shows a typical rescue. (H) Quantification of ngn1:GFP⁺ DRGs in control and gpr124 or reck morphants at 72 hpf after injection at the one-cell stage of 200 pg RNA encoding Gpr124 or Reck. ngn1:GFP⁺ DRGs were counted on one side of the larvae. (I) Lateral view of wild-type and reck morphants at 5 dpf. In all panels, values represent means ± SD (*p < 0.05; **p < 0.01; Kruskal–Wallis test). Morpholino and RNA injections were performed as described in ‘Methods’.