Figure 4
Figure 4. Vascular defects in VegfAb morphants. (A) Photomicrograph of a 5-bp mismatch vegfAb morpholino–injected (4.5 ng) fli1-gfp embryo at 2 dpf demonstrates no vascular defects (only one shown, although no phenotype was seen with either). (B) Defects in the formation of the ISVs anteriorly are clearly visible beginning at 2 dpf. (C) Photomicrograph of a 5-bp mismatch vegfAb control morpholino–injected (4.5 ng) fli1-gfp embryo at 4 dpf for comparison. (D-F) Beginning at 3 dpf, the number of circulating RBCs gradually decreased in 63% (162/255) of the embryos injected with 4.5 ng of the vegfAb ATG morpholino and in 47% (74/156) with 9 ng of the vegfAb-75 (5′UTR) morpholino (day-4 embryos are shown). Injection of an equivalent amount of 5-bp mismatch morpholinos did not affect vasculogenesis, and coinjection of the active vegfAb-75 (5′UTR) morpholino together with 50 pg each of vegfAb171 and vegfAb210 RNA reduced the number of embryos with defects in vasculogenesis by more than 50% (8/32 vs 22/40 abnormal embryo). VegfAb morphants showed decreased intersegmental vessel number and size (, ◀ in panels D and F) and aberrant head vascular development. SIVs were severely reduced in number, size, and branching that was more pronounced anteriorly. (E) At 4 dpf, blood is apparent in the head and anterior embryos (), which (F) corresponds to the areas with defects in angiogenesis. (G) Control morpholino–injected embryos demonstrate normal subintestinal vein (SIV) architecture by alkaline phosphatase staining, and (H) RBCs are shown by staining with o-dianisidine that stains hemoglobin reddish/brown. However, (I) injection of either morpholino targeting VegfAb leads to SIVs that are erratically placed and thin or nearly completely absent (only the start codon morpholino is shown) and (J) extravasation of RBCs in various structures, which is more pronounced anteriorly where the angiogenic defects are most visible (B-F). The results are combined from at least 3 separate experiments, and the photomicrographs are representative of the visible defects.

Vascular defects in VegfAb morphants. (A) Photomicrograph of a 5-bp mismatch vegfAb morpholino–injected (4.5 ng) fli1-gfp embryo at 2 dpf demonstrates no vascular defects (only one shown, although no phenotype was seen with either). (B) Defects in the formation of the ISVs anteriorly are clearly visible beginning at 2 dpf. (C) Photomicrograph of a 5-bp mismatch vegfAb control morpholino–injected (4.5 ng) fli1-gfp embryo at 4 dpf for comparison. (D-F) Beginning at 3 dpf, the number of circulating RBCs gradually decreased in 63% (162/255) of the embryos injected with 4.5 ng of the vegfAb ATG morpholino and in 47% (74/156) with 9 ng of the vegfAb-75 (5′UTR) morpholino (day-4 embryos are shown). Injection of an equivalent amount of 5-bp mismatch morpholinos did not affect vasculogenesis, and coinjection of the active vegfAb-75 (5′UTR) morpholino together with 50 pg each of vegfAb171 and vegfAb210 RNA reduced the number of embryos with defects in vasculogenesis by more than 50% (8/32 vs 22/40 abnormal embryo). VegfAb morphants showed decreased intersegmental vessel number and size (, ◀ in panels D and F) and aberrant head vascular development. SIVs were severely reduced in number, size, and branching that was more pronounced anteriorly. (E) At 4 dpf, blood is apparent in the head and anterior embryos (), which (F) corresponds to the areas with defects in angiogenesis. (G) Control morpholino–injected embryos demonstrate normal subintestinal vein (SIV) architecture by alkaline phosphatase staining, and (H) RBCs are shown by staining with o-dianisidine that stains hemoglobin reddish/brown. However, (I) injection of either morpholino targeting VegfAb leads to SIVs that are erratically placed and thin or nearly completely absent (only the start codon morpholino is shown) and (J) extravasation of RBCs in various structures, which is more pronounced anteriorly where the angiogenic defects are most visible (B-F). The results are combined from at least 3 separate experiments, and the photomicrographs are representative of the visible defects.

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal