Distribution and analysis of surface charge on brain endothelium in vitro and in situ

Vascular endothelial cells are associated with a number of anionic molecules. These anions are important in endothelial function, particularly in regulating permeability, haemostasis and cellular traffic. To explore the nature and distribution of anions on the brain endothelial cell (BEC) surface, we have examined rat brain endothelium in culture, and in situ. The anionic sites were probed with cationic colloidal gold and cationised ferritin, and visualised by light microscopy. Additionally we compared the distribution of the anionic sites on BEC with that present on other endothelial cell types in culture. The predominant anion detected on BEC was heparan sulphate (HS). This was distributed throughout the cell membrane, but was most densely associated with intercellular junctions. This pattern was distinct from the anionic locations observed in endothelia from aorta and epididymal fat microvessels. The distribution of anions was dependent on the age of cultured cells, with only minimal levels of HS seen at the periphery of younger cells. The nature and distribution of negative charge was different in situ. Here, sialic acid was the major surface anion, with only a small contribution from HS. The significance of these findings are discussed in relation to endothelial function in normal tissue and in pathological conditions.