Detection of glycosaminoglycans on the surface of human umbilical vein endothelial cells using gold-conjugated poly-L-lysine with silver enhancement.
Endothelial glycosaminoglycans are important in a diverse range of vascular functions. In the course of a biochemical and histological study exploring the role of glycosaminoglycans in inflammation, we have investigated the use of gold-conjugated poly-L-lysine with silver enhancement to establish the nature and physical location of glycosaminoglycans on the surface of cultured human umbilical vein endothelial cells. Cationic gold was effective in locating anionic sites in both cultured endothelial cells and in paraffin-embedded renal tissue. By manipulating pH, and by using enzymes specific for degrading glycosaminoglycans, it was found that, at pH 1.2, staining was directed primarily at glycosaminoglycans. The surface of human umbilical vein endothelial cells was found to be extensively covered in heparan sulphate, the histological appearance of which was dependent upon the fixation procedure employed. Heparan sulphate was also seen to co-distribute with the extracellular matrix protein, fibronectin, when endothelial cultures were simultaneously stained with cationic gold and an antibody to cellular fibronectin.
Klein
N J
NJ
Department of Paediatrics, St Mary's Hospital Medical School, London, UK.
Shennan
G I
GI
Heyderman
R S
RS
Levin
M
M
D001665:Binding Sites; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000737:chemistry* / Q000166:cytology; D005353:Fibronectins / Q000032:analysis*; D006025:Glycosaminoglycans / Q000032:analysis*; D006497:Heparan Sulfate / Q000032:analysis; D006651:Histocytochemistry; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007150:Immunohistochemistry; D007668:Kidney / Q000737:chemistry; D011107:Polylysine; D016622:Silver Staining; D014471:Umbilical Veins / Q000737:chemistry
D005353:Fibronectins; D006025:Glycosaminoglycans; D006497:Heparan Sulfate; D011107:Polylysine