A Zeiss LSM 700 confocal microscope was used for live imaging of the trunk and brain vasculature. Embryos were anaesthetized with a low dose of tricaine, placed in a glass-bottomed Petri dish (MatTek) on a layer of 1.2% low melt agarose and imaged using Plan-Apochromat 10×/0.45 and LCI Plan-Neofluar 25×/0.8 objectives.
In wt embryos, at 60 hpf, an average of 13 CtAs have connected to the basilar artery (BA)46 (link).
vegfaapromoter-less larvae were obtained from F2 heterozygous parents generated by outcrossing the founder and F1 fish.
To measure blood flow velocity, we performed time-lapse imaging of the trunk region of 78 hpf wild-type, maternal zygotic hbegfaΔ7 -/- and maternal zygotic hbegfafull locus del. -/- larvae using a Zeiss Spinning disc CSU-X1 confocal microscope with a high-speed camera, and quantified blood flow velocity in the dorsal aorta by measuring the time needed by erythrocytes to move 200 μm at the level of the fifth and sixth somites using Zen Blue as previously described47 (link).
hbegfafull locus del. larvae were obtained from F2 homozygous parents generated by outcrossing the founder.
hbegfaΔ7 and vegfaaΔ10 larvae were obtained from an incross of F3 heterozygous parents that were generated by consecutive outcrosses.
In wt embryos, at 60 hpf, an average of 13 CtAs have connected to the basilar artery (BA)46 (link).
vegfaapromoter-less larvae were obtained from F2 heterozygous parents generated by outcrossing the founder and F1 fish.
To measure blood flow velocity, we performed time-lapse imaging of the trunk region of 78 hpf wild-type, maternal zygotic hbegfaΔ7 -/- and maternal zygotic hbegfafull locus del. -/- larvae using a Zeiss Spinning disc CSU-X1 confocal microscope with a high-speed camera, and quantified blood flow velocity in the dorsal aorta by measuring the time needed by erythrocytes to move 200 μm at the level of the fifth and sixth somites using Zen Blue as previously described47 (link).
hbegfafull locus del. larvae were obtained from F2 homozygous parents generated by outcrossing the founder.
hbegfaΔ7 and vegfaaΔ10 larvae were obtained from an incross of F3 heterozygous parents that were generated by consecutive outcrosses.
