Vascular Basement Membranes: Preparation and Properties of Material Isolated with the Use of Detergents
During the past few years, work in our laboratories has been directed toward the isolation and chemical characterization of vascular basement membranes obtained from several organs. The chemical composition of these membranes, isolated in a pure form, can be related to changes in structure that occur in disease. We are particularly interested in those changes associated with diabetes mellitus (1–7). The nondisruptive detergent solubilization techniques used in these procedures permit the preparation of vascular basement membranes which are morphologically intact and which retain their histoarchitecture. With these preparations, the permeability of the basement membranes themselves, divorced from the contribution of the adjacent cell layers, was studied (3,5). Using this approach one can attempt to define the role which vascular basement membranes may play in the permeation of macromolecules through capillary walls, an area of some controversy (8–11). The findings that vascular basement membranes isolated by our techniques retained not only their morphologic and chemical properties (1–5), but also their immunologic characteristics (12), and their binding characteristics of cationic probes (13,14) further support the usefulness of this procedure in the study of basement membrane permeability. Modification of our technique has also been found useful in the isolation and examination of the pericellular matrix of human fibroblast cultures, a primary component of which is the cell adhesion protein fibronectin (15). In this chapter, we describe a method for isolation of vascular basement membranes, and compare permeability and chemical composition of membranes isolated from the central nervous system with those isolated from renal glomeruli. The method was first developed for study of renal glomeruli and these experiments will be described first.
KeywordsBasement Membrane Glomerular Basement Membrane Renal Glomerulus Pericellular Matrix Vascular Basement Membrane
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