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doi: 10.1242/10.1242/jcs.00600
Research Article |
1 Department of Pathology, Division of Molecular and Cellular Pathology and The
Cell Adhesion and Matrix Research Center, University of Alabama at Birmingham,
Birmingham, AL 35294-0019, USA
2 Research Service, Birmingham VA Medical Center, Birmingham, AL 35233-1996,
USA
3 Department of Pathology, Division of Laboratory Medicine, University of
Alabama at Birmingham, Birmingham, AL 35294-0019, USA
* Author for correspondence (e-mail: murphy{at}path.uab.edu)
Accepted 4 April 2003
The matricellular extracellular matrix protein thrombospondin-1 (TSP1) stimulates focal adhesion disassembly through a sequence (known as the hep I peptide) in its heparin-binding domain. This mediates signaling through a receptor co-complex involving calreticulin and low-density lipoprotein (LDL) receptor-related protein (LRP). We postulate that this transition to an intermediate adhesive state enhances cellular responses to dynamic environmental conditions. Since cell adhesion dynamics affect cell motility, we asked whether TSP1/hep I-induced intermediate adhesion alters cell migration. Using both transwell and Dunn chamber assays, we demonstrate that TSP1 and hep I gradients stimulate endothelial cell chemotaxis. Treatment with focal adhesion-labilizing concentrations of TSP1/hep I in the absence of a gradient enhances endothelial cell random migration, or chemokinesis, associated with an increase in cells migrating, migration speed, and total cellular displacement. Calreticulin-null and LRP-null fibroblasts do not migrate in response to TSP1/hep I, nor do endothelial cells treated with the LRP inhibitor receptor-associated protein (RAP). Furthermore, TSP1/hep I-induced focal adhesion disassembly is associated with reduced chemotaxis to basic fibroblast growth factor (bFGF) but enhanced chemotaxis to acidic (a)FGF, suggesting differential modulation of growth factor-induced migration. Thus, TSP1/hep I stimulation of intermediate adhesion regulates the migratory phenotype of endothelial cells and fibroblasts, suggesting a role for TSP1 in remodeling responses.
Key words: Thrombospondin-1, Focal adhesion, Cell migration, Calreticulin, LDL receptor-related protein
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