Fig. 5.
Fig. 5. Pattern of in situ hybridization of mRNA expression in human vascular tissue. In situ hybridization and immuno-histochemical staining was performed on serial sections (5 μm) of formaline-fixed and paraffin-embedded vascular tissue as described in Materials and Methods. Tissues were derived from the normal iliac artery of a 12-year-old male (A through D), 0.5 cm past the aortic bifurcation and the abdominal aorta of a 49-year-old woman (E through H), or a 39-year-old woman (I through T), both 1 cm before the bifurcation. (A) Masson Trichrome staining showing an overview through the iliac artery with the lumen on top, endothelial cells lining the vessel wall, the internal elastic lamina, the smooth muscle cell-containing media, and the spongy-appearing adventitia. (B) Specific staining of smooth muscle cells, using an antibody directed against SMC-specific -actin (1A4). (C) Autoradiographic image of the specific hybridization of an antisense probe for the endothelial cell specific protein von Willebrand factor, showing the integrity of endothelial cell mRNA and specificity of hybridization conditions. (D) Expression of novel cDNA CG12_1 is restricted to endothelial cells lining the vessel and the capillaries of the adventitia, identical to von Willebrand factor (C). (E) Masson Trichrome staining showing an overview through the aorta with the lumen on top, endothelial cells lining the vessel wall, a fibrous neo-intima (blue), the internal elastic lamina, the smooth muscle cell-containing media (purple), and the spongy-appearing adventitia. The boxed area is represented in (F) through (H). (F) Immunohistochemical staining using a monocyte/macrophage-specific antibody (HAM-56), showing the presence of extensive infiltration of the neointimal layer of the vessel wall, indicative of an inflammatory lesion. (G) Specific expression of MCP-1 by both macrophages and endothelial cells in this inflamed vessel wall. (H) Specific expression of ferritin by monocytes/macrophages and endothelial cells. (I) Masson Trichrome staining showing an overview through the aorta with the lumen on top, endothelial cells lining the vessel wall, a fibrous neo-intima (gray), the internal elastic lamina, the smooth muscle cell-containing media (blue/purple), and the spongy-appearing adventitia. The boxed area is represented in (J). (J) Immunostaining for macrophages (HAM-56) in this aorta section showing inflammation of the intima; the boxed area is represented in the in situ hybridizations shown in (K) through (N). (K) Expression pattern of MCP-1 in endothelial cells and macrophage/foam cells. (L) Specific expression of CA2_1 (hIAP-1) by the endothelial cells. (M) Expression of novel cDNA GG2_1 by endothelial cells. (M) and (N) are a darkfield representation of the autoradiographic images for greater clarity of hybridization of radiolabeled probes for sense mRNA for novel cDNA GG2_1 (M) and, as control for specificity, for antisense GG2_1 (N). (O) Masson Trichrome staining showing an overview through the aorta with the lumen on top, endothelial cells lining the vessel wall, a fibrous neo-intima (gray), the internal elastic lamina, the smooth muscle cell-containing media (blue/purple), and the spongy-appearing adventitia. (P) The integrity of the endothelial cell lining is shown with the specific lectin from Ulex europaeus, and specific expression of novel cDNA AG8_1 (stannin) is detected in endothelial cells (Q, darkfield representation). A more heavily inflamed area of the same aorta is shown by Masson Trichrome (R) and in the boxed area by immunostaining for macrophages (S) and expression of AG8_1 (T) in both endothelial cells and macrophages.

Pattern of in situ hybridization of mRNA expression in human vascular tissue. In situ hybridization and immuno-histochemical staining was performed on serial sections (5 μm) of formaline-fixed and paraffin-embedded vascular tissue as described in Materials and Methods. Tissues were derived from the normal iliac artery of a 12-year-old male (A through D), 0.5 cm past the aortic bifurcation and the abdominal aorta of a 49-year-old woman (E through H), or a 39-year-old woman (I through T), both 1 cm before the bifurcation. (A) Masson Trichrome staining showing an overview through the iliac artery with the lumen on top, endothelial cells lining the vessel wall, the internal elastic lamina, the smooth muscle cell-containing media, and the spongy-appearing adventitia. (B) Specific staining of smooth muscle cells, using an antibody directed against SMC-specific -actin (1A4). (C) Autoradiographic image of the specific hybridization of an antisense probe for the endothelial cell specific protein von Willebrand factor, showing the integrity of endothelial cell mRNA and specificity of hybridization conditions. (D) Expression of novel cDNA CG12_1 is restricted to endothelial cells lining the vessel and the capillaries of the adventitia, identical to von Willebrand factor (C). (E) Masson Trichrome staining showing an overview through the aorta with the lumen on top, endothelial cells lining the vessel wall, a fibrous neo-intima (blue), the internal elastic lamina, the smooth muscle cell-containing media (purple), and the spongy-appearing adventitia. The boxed area is represented in (F) through (H). (F) Immunohistochemical staining using a monocyte/macrophage-specific antibody (HAM-56), showing the presence of extensive infiltration of the neointimal layer of the vessel wall, indicative of an inflammatory lesion. (G) Specific expression of MCP-1 by both macrophages and endothelial cells in this inflamed vessel wall. (H) Specific expression of ferritin by monocytes/macrophages and endothelial cells. (I) Masson Trichrome staining showing an overview through the aorta with the lumen on top, endothelial cells lining the vessel wall, a fibrous neo-intima (gray), the internal elastic lamina, the smooth muscle cell-containing media (blue/purple), and the spongy-appearing adventitia. The boxed area is represented in (J). (J) Immunostaining for macrophages (HAM-56) in this aorta section showing inflammation of the intima; the boxed area is represented in the in situ hybridizations shown in (K) through (N). (K) Expression pattern of MCP-1 in endothelial cells and macrophage/foam cells. (L) Specific expression of CA2_1 (hIAP-1) by the endothelial cells. (M) Expression of novel cDNA GG2_1 by endothelial cells. (M) and (N) are a darkfield representation of the autoradiographic images for greater clarity of hybridization of radiolabeled probes for sense mRNA for novel cDNA GG2_1 (M) and, as control for specificity, for antisense GG2_1 (N). (O) Masson Trichrome staining showing an overview through the aorta with the lumen on top, endothelial cells lining the vessel wall, a fibrous neo-intima (gray), the internal elastic lamina, the smooth muscle cell-containing media (blue/purple), and the spongy-appearing adventitia. (P) The integrity of the endothelial cell lining is shown with the specific lectin from Ulex europaeus, and specific expression of novel cDNA AG8_1 (stannin) is detected in endothelial cells (Q, darkfield representation). A more heavily inflamed area of the same aorta is shown by Masson Trichrome (R) and in the boxed area by immunostaining for macrophages (S) and expression of AG8_1 (T) in both endothelial cells and macrophages.

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