Figure 7.

Endothelial-autonomous Vegfc and meningeal fibroblast-derived Vegfab and Vegfd are important for DLV formation.

(A–C) Dorsal views of 54 hpf vegfc+/+ (A), vegfcum18/+ (B), and vegfcum18/um18 (C) cranial vasculature visualized by Tg(kdrl:EGFP) expression. vegfcum18/um18 fish did not exhibit a defect in DLV formation (C) in contrast to the ‘No DLV’ phenotype found in vegfchu6410/hu6410 fish (Figure 1N). (D) Percentage of 54 hpf fish of indicated genotype with and without the DLV (n = 22 for vegfc+/+, n = 52 for vegfcum18/+, and n = 21 for vegfcum18/um18 fish). (E) Time course of the cell ablation experiments for panels (F–M). (F) Brightfield images of 72 hpf Tg(kdrl:EGFP), Tg(sox10:NTR), TgBAC(vegfab:NTR), and TgBAC(vegfd:NTR) larvae after treatment with DMSO or 10 mM Metronidazole (MTZ) from 34 to 72 hpf. (G–L) Dorsal views of 72 hpf Tg(kdrl:EGFP);TgBAC(vegfab:NTR) (G and H), Tg(kdrl:EGFP);TgBAC(vegfd:NTR) (I and J), and Tg(kdrl:EGFP);Tg(sox10:NTR) (K and L) cranial vasculature after treatment with DMSO (G, I, K) or MTZ (H, J, L) from 34 to 72 hpf. Efficient ablation of NTR-mCherry+ cells was observed after treatment with MTZ (H, J, L) compared to their respective DMSO-treated controls (G, I, K). MTZ-treated TgBAC(vegfab:NTR) and TgBAC(vegfd:NTR), but not Tg(sox10:NTR), larvae lacked the DLV (arrows). (M) Percentage of the indicated 72 hpf transgenic fish with and without the DLV after treatment with DMSO or MTZ (the number of the animals examined per each treatment group is listed in the graph). MTZ-treated TgBAC(vegfab:NTR) and TgBAC(vegfd:NTR) larvae exhibited a drastic increase in the ‘No DLV’ phenotype when compared to their corresponding DMSO treatment groups and the MTZ-treated Tg(kdrl:EGFP) larval group. In contrast, MTZ-treated Tg(kdrl:EGFP) and Tg(sox10:NTR) animals showed no significant differences compared to their corresponding DMSO treatment groups. Scale bars: 1 mm in (F); 50 μm in (C and L).

Expression Data

Expression Detail
Antibody Labeling
Phenotype Data

Phenotype Detail
Acknowledgments
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