Figure 2

Figure 2

GAH promotes zebrafish vascular development. (a,b) Vascular development status of transgenic fish with labeled vascular endothelial cells (red) and their nuclei (green) after treatment with different concentrations of GAH as indicated at 48 hpf using a laser scanning confocal microscope. Yellow arrows indicated: CtA, central arteries; PHBC, primordial hindbrain channel; MsV, mesencephalic vein; MtA, metencephalic artery; PHS, primary head sinus; VA, ventral aorta; AA, branchial arch; IOC, inner optic circle; OV, optic vein; PrA, prosencephalic artery; Se, intersegmental vessel; DLAV, dorsal longitudinal anastomotic vessel; CCV, common cardinal vein; CA, caudal artery; CV, caudal vein; PCeV, posterior (caudal) cerebral vein. Column 1 (vertical row) was a diagram of the blood vessels in the head region; Column 2 was a diagram of intersegmental vessels. Column 3 was a diagram of the nuclei of the vascular endothelial cells in the head; Column 4 was a diagram of the intersegmental vessels’ vascular endothelial cell nuclei. The experiment was repeated two times. (c–e) Statistical data of (a,b). (f) Expression of kdrl at 24 hpf and 48 hpf by ISH with increasing concentrations of GAH as indicated. Red arrows indicated MCeV, middle cerebral vein; PMBC, primordial midbrain channel; DA, dorsal aorta; CA, caudal artery; PCV, posterior (caudal) cardinal vein; CV, caudal vein; Se, intersegmental vessel. (g) Statistical data of (f) based on three (24 hpf) and four (48 hpf) repetitions. (h) BrdU incorporation assay to investigate cell proliferation activity. Green fluorescence indicated a vascular endothelial nucleus; red fluorescence indicated BrdU-positive cells. The experiment was repeated twice. Scale bars: 100 μm. (i) Statistical data of the number of BrdU-positive cells co-located with GFP in (h). p < 0.05 (*), p < 0.01 (**), and p < 0.001 (***); NS, nonsignificant.