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Expression sites of colligin 2 in glioma blood vessels.

Mustafa D, van der Weiden M, Zheng P, Nigg A, Luider TM, Kros JM - Brain Pathol. (2009)

Bottom Line: Expression of colligin 2 was also found in cells scattered around blood vessels and in few glial fibrillary acidic protein-positive cells within the blood vessels.There is overlap in the expression of colligin 2 and the collagens type I and IV for which colligin 2 is a chaperon.We conclude that colligin 2 is expressed in all cellular components of glioma blood vessels and may serve as a general marker for active angiogenesis.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology and Laboratory of Neuro-oncology and Clinical Proteomics, Erasmus Medical Center, Dr. Molewaterplein 50, 3015 GD Rotterdam, The Netherlands.

ABSTRACT
In a previous study using state-of-the-art proteomic techniques, we identified colligin 2 (HSP47) as a glioma blood vessel-specific protein. In the present study we precisely localized the expression of colligin 2 in the blood vessels of diffusely infiltrating gliomas and relate the expression to the distinct cellular components of the vessels by using multiple immunolabeling and confocal microscopy. We grouped the glioma blood vessels into morphological categories ranging from normal looking capillaries to vessels with hypertrophic and sclerotic changes. The expression patterns of various markers of endothelial and pericytic differentiation were correlated with the position of the cells in the vessels and the expression of colligin 2. We found that colligin 2 is expressed in all categories of glioma blood vessels in cells with endothelial and pericytic lineage. Expression of colligin 2 was also found in cells scattered around blood vessels and in few glial fibrillary acidic protein-positive cells within the blood vessels. There is overlap in the expression of colligin 2 and the collagens type I and IV for which colligin 2 is a chaperon. We conclude that colligin 2 is expressed in all cellular components of glioma blood vessels and may serve as a general marker for active angiogenesis.

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Expression of the various markers in layered hypertrophic blood vessels in glioma. CD31 and CD34 are expressed in the endothelial layer. CD105 and endosialin are expressed in the endothelium and the external layer. αSMA, NG2 and collagen IV are expressed in the intermediate and external layers, not in the endothelium. Colligin 2 is expressed in all layers of the vessels (panels A–F: ×40). G–K. Confocal images of layered hypertrophic blood vessels in glioma for the expression of CD105. Double immunolabeling for CD105/colligin 2. The endothelial cells and the external cells express both CD105 and colligin 2, while the intermediate cells express colligin 2 only (arrows) (panel H: green = CD105; panel I: red = colligin 2; panel J: blue = DAPI; panel K: merged picture).
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fig03: Expression of the various markers in layered hypertrophic blood vessels in glioma. CD31 and CD34 are expressed in the endothelial layer. CD105 and endosialin are expressed in the endothelium and the external layer. αSMA, NG2 and collagen IV are expressed in the intermediate and external layers, not in the endothelium. Colligin 2 is expressed in all layers of the vessels (panels A–F: ×40). G–K. Confocal images of layered hypertrophic blood vessels in glioma for the expression of CD105. Double immunolabeling for CD105/colligin 2. The endothelial cells and the external cells express both CD105 and colligin 2, while the intermediate cells express colligin 2 only (arrows) (panel H: green = CD105; panel I: red = colligin 2; panel J: blue = DAPI; panel K: merged picture).

Mentions: The vessels encountered in the glioma samples were divided in: small vessels including capillaries, which did not show morphological changes; vessels with hypertrophied walls (either with organized layering as in normal larger blood vessels or with disorganized, haphazardly arranged cellular components); vessels with glomeruloid appearance and vessels with signs of recanalization. The lumina-lining cells of capillaries in glioma were invariably positive for CD31, CD34 and CD105 (Figure 2; Table 4). In addition, there was immunopositivity for αSMA, endosialin and NG2. Expression of colligin 2 was found in all small blood vessels (Table 4). In the vessels with hypertrophied walls with organized layering, the endothelium expressed CD31 and CD34 while the intermediate and the external layers expressed αSMA and NG2 (Table 4). The inner (endothelial) and outer layer of these vessels expressed endosialin and CD105 while cells in between these two layers remained negative for these two markers. Colligin 2 was expressed in all cell layers of the hypertrophied blood vessels (Figure 3). In the hypertrophied vessels with disorganized arranged cellular components, the internal diameter of the lumen appeared irregular while the external diameter of the vessel wall varied per segment of the vessel. The endothelium stained positive for CD31, CD34 and CD105 but not for αSMA, NG2 or endosialin (Table 4). The abluminal cells of these vessels stained positive for CD105, αSMA, NG2 and endosialin. Colligin 2 was expressed in all layers of the vessel walls (Figure 4). In the glomeruloid blood vessels, the cells are haphazardly arranged around multiple small lumina. The lumina-lining cells of the glomeruloid blood vessels were positive for CD31, CD34 and CD105. The cells in the abluminal position stained positive for endosialin, NG2, and αSMA (Table 4). The expression of colligin 2 was positive in all cellular components of these blood vessels (Figure 5). The lumina-lining cells of the larger recanalized vessels were positive for CD31, CD34 and CD105 (Table 4). The other cells stained positive for αSMA, endosialin and colligin 2. Remarkably, a minority of cells in the recanalized thrombi remained negative for all markers used (Figure 6).


Expression sites of colligin 2 in glioma blood vessels.

Mustafa D, van der Weiden M, Zheng P, Nigg A, Luider TM, Kros JM - Brain Pathol. (2009)

Expression of the various markers in layered hypertrophic blood vessels in glioma. CD31 and CD34 are expressed in the endothelial layer. CD105 and endosialin are expressed in the endothelium and the external layer. αSMA, NG2 and collagen IV are expressed in the intermediate and external layers, not in the endothelium. Colligin 2 is expressed in all layers of the vessels (panels A–F: ×40). G–K. Confocal images of layered hypertrophic blood vessels in glioma for the expression of CD105. Double immunolabeling for CD105/colligin 2. The endothelial cells and the external cells express both CD105 and colligin 2, while the intermediate cells express colligin 2 only (arrows) (panel H: green = CD105; panel I: red = colligin 2; panel J: blue = DAPI; panel K: merged picture).
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2805918&req=5

fig03: Expression of the various markers in layered hypertrophic blood vessels in glioma. CD31 and CD34 are expressed in the endothelial layer. CD105 and endosialin are expressed in the endothelium and the external layer. αSMA, NG2 and collagen IV are expressed in the intermediate and external layers, not in the endothelium. Colligin 2 is expressed in all layers of the vessels (panels A–F: ×40). G–K. Confocal images of layered hypertrophic blood vessels in glioma for the expression of CD105. Double immunolabeling for CD105/colligin 2. The endothelial cells and the external cells express both CD105 and colligin 2, while the intermediate cells express colligin 2 only (arrows) (panel H: green = CD105; panel I: red = colligin 2; panel J: blue = DAPI; panel K: merged picture).
Mentions: The vessels encountered in the glioma samples were divided in: small vessels including capillaries, which did not show morphological changes; vessels with hypertrophied walls (either with organized layering as in normal larger blood vessels or with disorganized, haphazardly arranged cellular components); vessels with glomeruloid appearance and vessels with signs of recanalization. The lumina-lining cells of capillaries in glioma were invariably positive for CD31, CD34 and CD105 (Figure 2; Table 4). In addition, there was immunopositivity for αSMA, endosialin and NG2. Expression of colligin 2 was found in all small blood vessels (Table 4). In the vessels with hypertrophied walls with organized layering, the endothelium expressed CD31 and CD34 while the intermediate and the external layers expressed αSMA and NG2 (Table 4). The inner (endothelial) and outer layer of these vessels expressed endosialin and CD105 while cells in between these two layers remained negative for these two markers. Colligin 2 was expressed in all cell layers of the hypertrophied blood vessels (Figure 3). In the hypertrophied vessels with disorganized arranged cellular components, the internal diameter of the lumen appeared irregular while the external diameter of the vessel wall varied per segment of the vessel. The endothelium stained positive for CD31, CD34 and CD105 but not for αSMA, NG2 or endosialin (Table 4). The abluminal cells of these vessels stained positive for CD105, αSMA, NG2 and endosialin. Colligin 2 was expressed in all layers of the vessel walls (Figure 4). In the glomeruloid blood vessels, the cells are haphazardly arranged around multiple small lumina. The lumina-lining cells of the glomeruloid blood vessels were positive for CD31, CD34 and CD105. The cells in the abluminal position stained positive for endosialin, NG2, and αSMA (Table 4). The expression of colligin 2 was positive in all cellular components of these blood vessels (Figure 5). The lumina-lining cells of the larger recanalized vessels were positive for CD31, CD34 and CD105 (Table 4). The other cells stained positive for αSMA, endosialin and colligin 2. Remarkably, a minority of cells in the recanalized thrombi remained negative for all markers used (Figure 6).

Bottom Line: Expression of colligin 2 was also found in cells scattered around blood vessels and in few glial fibrillary acidic protein-positive cells within the blood vessels.There is overlap in the expression of colligin 2 and the collagens type I and IV for which colligin 2 is a chaperon.We conclude that colligin 2 is expressed in all cellular components of glioma blood vessels and may serve as a general marker for active angiogenesis.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology and Laboratory of Neuro-oncology and Clinical Proteomics, Erasmus Medical Center, Dr. Molewaterplein 50, 3015 GD Rotterdam, The Netherlands.

ABSTRACT
In a previous study using state-of-the-art proteomic techniques, we identified colligin 2 (HSP47) as a glioma blood vessel-specific protein. In the present study we precisely localized the expression of colligin 2 in the blood vessels of diffusely infiltrating gliomas and relate the expression to the distinct cellular components of the vessels by using multiple immunolabeling and confocal microscopy. We grouped the glioma blood vessels into morphological categories ranging from normal looking capillaries to vessels with hypertrophic and sclerotic changes. The expression patterns of various markers of endothelial and pericytic differentiation were correlated with the position of the cells in the vessels and the expression of colligin 2. We found that colligin 2 is expressed in all categories of glioma blood vessels in cells with endothelial and pericytic lineage. Expression of colligin 2 was also found in cells scattered around blood vessels and in few glial fibrillary acidic protein-positive cells within the blood vessels. There is overlap in the expression of colligin 2 and the collagens type I and IV for which colligin 2 is a chaperon. We conclude that colligin 2 is expressed in all cellular components of glioma blood vessels and may serve as a general marker for active angiogenesis.

Show MeSH
Related in: MedlinePlus