The build-up of the thrombospondin extracellular matrix. An apparent dependence on synthesis and on preformed fibrillar matrix.
Thrombospondin (TSP) is a multifunctional protein synthesized by several cell types in culture, among them endothelial cells, and incorporated into the extracellular matrix (ECM) of these cells. In vitro it has been detected in most interstitial spaces, and its presence has been suggested to be developmentally controlled. With the aim of elucidating the role of TSP in the extracellular matrix, we studied the build-up of this protein in the matrix as a function of cell type density and age in culture. The development of the TSP matrix was compared to that of fibronectin (FN) and of von Willebrand Factor (VWF) in the same cultures using our monoclonal anti-TSP antibody B7-3 and monospecific anti-FN or anti-VWF antibodies. In cultures of bovine aortic endothelial cells (BAEC) we observed that both the pattern and distribution of TSP in the matrix changed with cell density and time in culture; it started as a completely amorphous dense layer of protein when the cells were sparse and changed to well organized fibrils when the cells had been confluent for a while. The first appearance of the fibrillar arrays of TSP coincides perfectly with that of the FN fibers; extracellular VWF also was first detected at this point. From this time in culture, there was an increasing coincidence of the TSP with the FN and the VWF matrices suggesting the formation of a complex multicomponent structure of the fibrillar network.(ABSTRACT TRUNCATED AT 250 WORDS)
Dreyfus
M
M
Department of Polymer Research, Weizmann Institute of Science, Rehovot, Israel.
Lahav
J
J
D000375:Aging / Q000378:metabolism; D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology / Q000648:ultrastructure; D005109:Extracellular Matrix / Q000032:analysis / Q000378:metabolism* / Q000502:physiology; D005353:Fibronectins / Q000032:analysis; D005455:Fluorescent Antibody Technique; D006023:Glycoproteins / Q000378:metabolism*; D019699:Thrombospondins; D014841:von Willebrand Factor / Q000032:analysis
D005353:Fibronectins; D006023:Glycoproteins; D019699:Thrombospondins; D014841:von Willebrand Factor