Loss of <i>sema3fb</i> angiogenic deficits are independent of blood flow.

Abstract

A) Confocal lateral images of laminin-stained embryos at 30hpf. Tg(kdrl:mCherry) endothelium (red) and laminin (green). Embryos derived from a heterozygous sema3fbca305/+ incross. B) Quantification of the length of ISA sprouts at 30hpf, N = 1: wild type (WT) (40 ISAs, 4 embryos, mean of 102±1 μm2), sema3fbca305/+ (120 ISAs, 12 embryos, mean of 90±8 μm2), and sema3fbca305 (150 ISAs, 15 embryos, mean of 90±14 μm2). C) Confocal lateral images of the trunk endothelium (black) in blood flow-stopped tnnt2ATG-MO injected wild type siblings (WT) and sema3fbca305 mutants. DLAV gaps (blue asterisks) and truncated ISAs sprouts (yellow arrowheads) are marked. Scale bar, 100μm. D) Length of ISA sprouts at 30 hpf, N = 3: WT (80 ISAs, 8 embryos, mean length of 106±3 μm), WT + tnnt2aATG-MO (100 ISAs, 10 embryos, mean 98±8.4 μm), sema3fbca305 (80 ISAs, 8 embryos, mean 86±17 μm), and sema3fbca305 + tnnt2aATG-MO (100 ISAs, 10 embryos, mean 85±22 μm). E) Percentage of ISAs connected at DLAV at 30 hpf, N = 3: WT (mean 82±9% connected), WT + tnnt2aATG-MO (mean 86±7% connected), sema3fbca305 (80 ISAs, 8 embryos, mean 54±15% connected), and sema3fbca305 + tnnt2aATG-MO (mean 54±16% connected). F) Quantification of width of DA in 30 hpf embryos, N = 3: WT (8 embryos, 5 measurements per embryo/n = 40 total, mean width of 18±3 μm), WT + tnnt2aATG-MO (10 embryos, 5 measurements per embryo/n = 50 total, mean 9±1 μm), sema3fbca305 (8 embryos, 5 measurements per embryo/n = 40 total, mean 11±3 μm), and sema3fbca305 + tnnt2aATG-MO (10 embryos, 5 measurements per embryo/n = 50 total, mean 10±1 μm). G) Quantification of width of PCV in 30 hpf embryos, N = 3: WT (n = 40, mean width of 22±3 μm), WT + tnnt2aATG-MO (n = 50, mean 19±4 μm), sema3fbca305 (n = 40, mean 20±3 μm), and sema3fbca305 + tnnt2aATG-MO(n = 50, mean 19±4 μm). 2-Way ANOVA Tukey’s multiple comparisons test, **** means p = (TIF)</p