Fig. 4.
Fig. 4. Abnormal vascular morphology in CAGp-ephrin-B2 Tg mice. / (A) Western blotting of extracts (20 μg protein) from the spleens of neonatal ephrin-B2 Tg mice with anti–ephrin-B2 polyclonal antibody. Arrowhead indicates overexpressed ephrin-B2 protein in CAGp-ephrin-B2 Tg mice. (B) Vascular phenotypes of wild-type (left) and CAGp-ephrin-B2Tg/Tg mice (right) at neonatal stages. The dilated aorta is observed, especially in the ascending portion (stars). (C-F) Cross sections of the ascending aorta stained with hematoxylin and eosin were examined after the aortic dissection event. Tg/Tg mice (D, F) with aortic dissection (arrowheads in F). Bars indicate 100 μm. (G-J) Light and electron micrographs comparing wild-type (G, I) andTg/Tg mouse embryos (H, J) at E19. Sections were stained with toluidine blue. Arrowheads in panel H show a thin vascular wall. Bars indicate 150 μm (G, H). Electron micrographs of the same vessel (G, H) at magnification × 1700 (I, J). In electron micrographs, SMCs (stars) and elastic bands (arrows) have shown the complementary arrangement pattern in wild-type mice (I). In contrast, they have disappeared, and new microvessels (arrows) are observed in this space (J). ECs in Tg/Tg mice have a flat morphology with a budlike structure (arrowheads in J), while ECs in wild-type mice show a round morphology (arrowheads in I). (K-M) Explant culture of ascending aorta from a CAGp-ephrin-B2 Tg/Tg mouse (L) and a wild-type littermate (K) were examined by staining with the anti–smooth muscle actin antibody. Bars indicate 1 mm. (M) Quantitative analysis of the area of smooth muscle actin+ cells was performed with the NIH image computer analyzing system. Each column represents the mean area, and the error bars indicate SD (n = 8). (N) In vitro proliferation analysis. Identical numbers of cells of each genotype were plated. The growth of these cultures was measured by counting the total number of cells on each of the indicated days. A representative experiment is shown. Similar results were obtained from SMCs derived from all 3 genotypes.

Abnormal vascular morphology in CAGp-ephrin-B2 Tg mice.

(A) Western blotting of extracts (20 μg protein) from the spleens of neonatal ephrin-B2 Tg mice with anti–ephrin-B2 polyclonal antibody. Arrowhead indicates overexpressed ephrin-B2 protein in CAGp-ephrin-B2 Tg mice. (B) Vascular phenotypes of wild-type (left) and CAGp-ephrin-B2Tg/Tg mice (right) at neonatal stages. The dilated aorta is observed, especially in the ascending portion (stars). (C-F) Cross sections of the ascending aorta stained with hematoxylin and eosin were examined after the aortic dissection event. Tg/Tg mice (D, F) with aortic dissection (arrowheads in F). Bars indicate 100 μm. (G-J) Light and electron micrographs comparing wild-type (G, I) andTg/Tg mouse embryos (H, J) at E19. Sections were stained with toluidine blue. Arrowheads in panel H show a thin vascular wall. Bars indicate 150 μm (G, H). Electron micrographs of the same vessel (G, H) at magnification × 1700 (I, J). In electron micrographs, SMCs (stars) and elastic bands (arrows) have shown the complementary arrangement pattern in wild-type mice (I). In contrast, they have disappeared, and new microvessels (arrows) are observed in this space (J). ECs in Tg/Tg mice have a flat morphology with a budlike structure (arrowheads in J), while ECs in wild-type mice show a round morphology (arrowheads in I). (K-M) Explant culture of ascending aorta from a CAGp-ephrin-B2 Tg/Tg mouse (L) and a wild-type littermate (K) were examined by staining with the anti–smooth muscle actin antibody. Bars indicate 1 mm. (M) Quantitative analysis of the area of smooth muscle actin+ cells was performed with the NIH image computer analyzing system. Each column represents the mean area, and the error bars indicate SD (n = 8). (N) In vitro proliferation analysis. Identical numbers of cells of each genotype were plated. The growth of these cultures was measured by counting the total number of cells on each of the indicated days. A representative experiment is shown. Similar results were obtained from SMCs derived from all 3 genotypes.

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