Figure 1.
Figure 1. syndecan-2 MO–injected embryos showed angiogenic defects. (A) An uninjected wild-type embryo at 48 hpf. (B) Microangiogram of an uninjected wild-type embryo at 48 hpf. Bright field image (C) and microangiogram (D) of an embryo injected with 5 ng of a 4-base mismatch syndecan-2 showing intact vasculature. (E) An embryo injected with 5 ng syndecan-2 MO1, showing no obvious morphologic defects at 48 hpf. (F,I,J) Microangiograms of embryos injected with 5 ng syndecan-2 MO1, showing vascular phenotypes (frequency: 84% ± 4%, n = 55). Magnified images of sections indicated by brackets in panels B and F are shown in panels G and H, respectively. Embryos injected with syndecan-2 MO1 showed either aberrant (F; arrowheads in H compared with wild-type in G) or reduced intersegmental vessels (I, arrows) and reduced tail vessels (I, arrowheads). In the most severe case, embryos showed a complete lack of intersegmental vessels (top arrows in J) and lack of tail vessels (bottom arrows in J). In situ analysis was also performed to assess molecular phenotypes of syndecan-2 MO–injected embryos. (K-L) Expression of ephrin-B2 in a wild-type embryo (K) and a syndecan-2 MO–injected embryo (L) at 24 hpf. Arterial expression of ephrin-B2 was retained in syndecan-2 MO–injected embryos. (M-N) Expression of VEGF-A in wild-type (M) and syndecan-2 MO–injected (N) embryos (n = 20). Somitic expression of VEGF-A was not affected in syndecan-2 MO–injected embryos. Cross sections from the anterior trunk of a wild-type embryo (O) and a syndecan-2 MO–injected embryo (P) at 44 to 48 hpf stained with hematoxylin and eosin. The dorsal aorta (top arrowhead) and the axial vein (bottom arrowhead) were both observed in syndecan-2 MO–injected embryos. (Q) Expression of fli-1 in a wild-type embryo at 24 hpf. (R) A syndecan-2 MO–injected embryo at 24 hpf, showing a lack of intersegmental fli-1 expression (arrowheads). Of the embryos, 58% (n = 35) showed reduced intersegmental fli-1 expression at 5 ng syndecan-2 MO1. (S) Expression of flk-1 in a wild-type embryo at 24 hpf. (T) A syndecan-2 MO–injected embryo at 24 hpf, showing a lack of intersegmental flk-1 expression (arrowheads). Of embryos, 50% (n = 40) showed reduced flk-1 expression at 5 ng syndecan-2 MO1. (U) Expression of tie-1 in a wild-type embryo at 24 hpf. (V) A syndecan-2 MO–injected embryo at 24 hpf, showing a lack of intersegmental expression of tie-1 (arrowheads). At 5 ng syndecan-2 MO1, 63% of embryos (n = 56) showed reduced tie-1 expression. (A,C,E) Bright field images (original magnification, × 5). (B,D,F-J) Microangiography analysis (original magnification, × 5. (K-N,Q-V) In situ hybridization (original magnification, × 10). (O-P) Original magnification, × 40.

syndecan-2 MO–injected embryos showed angiogenic defects. (A) An uninjected wild-type embryo at 48 hpf. (B) Microangiogram of an uninjected wild-type embryo at 48 hpf. Bright field image (C) and microangiogram (D) of an embryo injected with 5 ng of a 4-base mismatch syndecan-2 showing intact vasculature. (E) An embryo injected with 5 ng syndecan-2 MO1, showing no obvious morphologic defects at 48 hpf. (F,I,J) Microangiograms of embryos injected with 5 ng syndecan-2 MO1, showing vascular phenotypes (frequency: 84% ± 4%, n = 55). Magnified images of sections indicated by brackets in panels B and F are shown in panels G and H, respectively. Embryos injected with syndecan-2 MO1 showed either aberrant (F; arrowheads in H compared with wild-type in G) or reduced intersegmental vessels (I, arrows) and reduced tail vessels (I, arrowheads). In the most severe case, embryos showed a complete lack of intersegmental vessels (top arrows in J) and lack of tail vessels (bottom arrows in J). In situ analysis was also performed to assess molecular phenotypes of syndecan-2 MO–injected embryos. (K-L) Expression of ephrin-B2 in a wild-type embryo (K) and a syndecan-2 MO–injected embryo (L) at 24 hpf. Arterial expression of ephrin-B2 was retained in syndecan-2 MO–injected embryos. (M-N) Expression of VEGF-A in wild-type (M) and syndecan-2 MO–injected (N) embryos (n = 20). Somitic expression of VEGF-A was not affected in syndecan-2 MO–injected embryos. Cross sections from the anterior trunk of a wild-type embryo (O) and a syndecan-2 MO–injected embryo (P) at 44 to 48 hpf stained with hematoxylin and eosin. The dorsal aorta (top arrowhead) and the axial vein (bottom arrowhead) were both observed in syndecan-2 MO–injected embryos. (Q) Expression of fli-1 in a wild-type embryo at 24 hpf. (R) A syndecan-2 MO–injected embryo at 24 hpf, showing a lack of intersegmental fli-1 expression (arrowheads). Of the embryos, 58% (n = 35) showed reduced intersegmental fli-1 expression at 5 ng syndecan-2 MO1. (S) Expression of flk-1 in a wild-type embryo at 24 hpf. (T) A syndecan-2 MO–injected embryo at 24 hpf, showing a lack of intersegmental flk-1 expression (arrowheads). Of embryos, 50% (n = 40) showed reduced flk-1 expression at 5 ng syndecan-2 MO1. (U) Expression of tie-1 in a wild-type embryo at 24 hpf. (V) A syndecan-2 MO–injected embryo at 24 hpf, showing a lack of intersegmental expression of tie-1 (arrowheads). At 5 ng syndecan-2 MO1, 63% of embryos (n = 56) showed reduced tie-1 expression. (A,C,E) Bright field images (original magnification, × 5). (B,D,F-J) Microangiography analysis (original magnification, × 5. (K-N,Q-V) In situ hybridization (original magnification, × 10). (O-P) Original magnification, × 40.

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