Molecular characterization of anionic sites on the luminal front of endoneurial capillaries in sciatic nerve.
The distribution of anionic microdomains has been described in cerebral vessels and more recently in capillaries of peripheral nerve. Evidence is accumulating that these sites play a role in the barrier function of vascular endothelia in the PNS and CNS. The chemical nature of anionic sites has been at least partly determined for cerebral vessels but not in peripheral nerve. This study reports our preliminary investigations to determine the nature of endothelial anionic sites in sciatic nerve. The effects of digestion of ultra-thin sections of nerve with a battery of proteolytic and glycolytic enzymes (papain, trypsin, proteinase K, hyaluronidase, heparinase, heparitinase and neuraminidase) on the distribution of anionic sites was determined using the label, cationic colloidal gold. Papain, a proteolytic enzyme of broad specificity, succeeded in removing the majority of cationic colloidal gold-binding sites on the luminal surface of vascular endothelia. In contrast trypsin and proteinase K were less effective, reflecting their narrower specificity. Hyaluronidase, heparinase and heparitinase did not significantly affect cationic colloidal gold-labelling. However, a considerable reduction in cationic colloidal gold-binding occurred following neuraminidase digestion. These results suggest that, as in cerebral vessels, sialic acid-containing glycoproteins are largely responsible for the negatively charged domains on the luminal membrane of endothelial cells in peripheral nerve.
Lawrenson
J G
JG
Reta Lila Weston Institute of Neurological Studies, University College London Medical School, UK.
Reid
A R
AR
Allt
G
G
D000818:Animals; D000838:Anions / Q000737:chemistry; D002196:Capillaries / Q000502:physiology; D006019:Glycolysis / Q000502:physiology; D017979:Gold Colloid; D008297:Male; D010447:Peptide Hydrolases; D011107:Polylysine; D051381:Rats; D017207:Rats, Sprague-Dawley; D012584:Sciatic Nerve / Q000098:blood supply*
D000838:Anions; D017979:Gold Colloid; D011107:Polylysine; D010447:Peptide Hydrolases