Partial chemical characterization of the anionic sites in the basal lamina of fenestrated capillaries.
The distribution of anionic sites in the basal laminae of the blood capillaries of the murine pancreas was studied in specimens fixed in ruthenium red (RR)-glutaraldehyde mixtures. The sites appeared as discrete, small (6 to 18 nm) particles distributed throughout the three laminae but concentrated primarily in the lamina rara externa, in which--spaced 80-100 nm apart--they formed a planar, partially ordered lattice comparable to that revealed by cationized ferritin in previous studies (M. Simionescu, N. Simionescu, and G. E. Palade, 1982, J. Cell Biol. 95, 425-434). The chemical nature of the anionic sites was explored by incubating fresh tissue specimens in solutions of selected enzymes before fixation in RR-glutaraldehyde mixtures. Pronase P and papain removed completely the anionic sites and left behind an extensively degraded and disorganized basal lamina. Trypsin caused the removal of anionic sites only, did not degrade the rest of the basal lamina, but detached it completely from the endothelium. Chondroitinase ABC reduced slightly the size and the surface density of RR-stainable particles, and detached focally the rest of the basal lamina from the endothelium and pericytes. Crude heparinase caused a nearly complete removal of anionic sites, and pure heparitinase gave comparable but less extensive results. Similar effects were recorded on the basal laminae of smooth muscle fibers and pancreatic acini and ducts. The results indicate that the anionic sites of all basal laminae examined are contributed primarily by heparin sulfate proteoglycans and trace amounts of chondroitin sulfate proteoglycans.
D000818:Animals; D000838:Anions / Q000032:analysis*; D001485:Basement Membrane / Q000032:analysis*; D002196:Capillaries / Q000032:analysis; D002810:Chondroitinases and Chondroitin Lyases; D051379:Mice; D009439:Neuraminidase; D010179:Pancreas / Q000098:blood supply*; D010206:Papain; D011133:Polysaccharide-Lyases; D011402:Pronase; D014357:Trypsin
D000838:Anions; D009439:Neuraminidase; D014357:Trypsin; D010206:Papain; D011402:Pronase; D002810:Chondroitinases and Chondroitin Lyases; D011133:Polysaccharide-Lyases; C021349:heparitinsulfate lyase