Three-dimensional organization of the extracellular matrix secreted by cultured rat smooth muscle cells.
Specific interactions between cells and the extracellular matrix (ECM) in which they are embedded play a vital role in tissue organization. In recent years, many of the individual components of the extracellular matrix have been isolated and their molecular structures elucidated, but the detailed topography of most extracellular matrices, as they are deposited by cells, is still largely unknown. In this study, the insoluble extracellular matrix produced by cultured rat vascular smooth muscle cells has been characterized morphologically using high-resolution electron microscopy of rotary platinum replicas. These cells grew as flat sheets in culture, secreting their matrix laterally and basally. The matrix was composed of a cross-linked fibrillar meshwork. Some fine fibers (10 to 15 nm in diameter) were naked, but most of the filamentous mesh was covered with coarse granular material. Limited digestion with trypsin or pancreatic elastase removed most of this coating, indicating that the granules were glycoproteins and proteoglycans. Another subset of matrix fibrils (20 to 40 nm in diameter) was identified as type I collagen by direct comparison with purified bovine skin collagen. In addition to exposing the underlying filamentous substructure of the matrix, protease treatment also revealed large, straight fiber bundles and globules of amorphous material suspended in the filamentous web. This novel view of a complex matrix promises to provide spatial information that will be useful in future studies of cell interactions with the ECM.
Aggeler
J
J
Department of Human Anatomy, University of California, Davis 95616.
D064753:Ammonium Hydroxide; D000818:Animals; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism; D004549:Elastin / Q000378:metabolism; D005109:Extracellular Matrix / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure*; D006023:Glycoproteins / Q000378:metabolism; D006878:Hydroxides / Q000494:pharmacology; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning; D009131:Muscle, Smooth, Vascular / Q000648:ultrastructure*; D010196:Pancreatic Elastase / Q000378:metabolism; D051381:Rats; D014357:Trypsin / Q000378:metabolism
D006023:Glycoproteins; D006878:Hydroxides; D064753:Ammonium Hydroxide; D003094:Collagen; D004549:Elastin; D010196:Pancreatic Elastase; D014357:Trypsin