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Vol. 8, Issue 11, 2345-2363, November 1997
Medical Research Council Laboratory for Molecular Cell Biology and
Department of Biochemistry and Molecular Biology, University College
London, London WC1E 6BT, United Kingdom
Cell adhesion to thrombospondin-1 (TSP-1) correlates with assembly
of cell-substratum contact structures that contain fascin microspikes.
In this analysis, cell-matrix requirements for assembly of fascin
microspikes were examined in detail. In six cell lines, cell spreading
on a TSP-1 substratum correlated with expression of fascin protein and
formation of fascin microspikes. Microspikes were not formed by H9c2
cells adherent on fibronectin, vitronectin, collagen IV, or platelet
factor 4. However, both fascin microspikes and focal contacts were
assembled by cells adherent on laminin-1. Using mixed substrata
containing different proportions of TSP-1, and fibronectin, fascin
microspike formation by H9c2 and C2C12 cells was found to be reduced on
substrata containing 25% fibronectin and abolished on substrata
containing 75% fibronectin. Adhesion to intermediate mixtures of TSP-1
and fibronectin resulted in coassembly of fascin microspikes and focal
contacts, colocalization of fascin with actin stress fiber bundles and
altered distributions of 
1 integrins, cortical 
-actinin,
and tropomyosin. In cells adherent on 50% TSP-1:50% fibronectin,
GRGDSP peptide treatment decreased focal contact assembly and altered
cytoskeletal organization but did not inhibit microspike assembly.
Treatment with chondroitin sulfate A or p-nitrophenol
-D-xylopyranoside decreased microspike formation and
modified cytoskeletal organization but did not inhibit focal contact
formation. In polarized migratory and postmitotic C2C12 cells, fascin
microspikes and ruffles were localized at leading edges and TSP matrix
deposition was also concentrated in this region. Depletion of matrix
TSP by heparin treatment correlated with decreased microspike formation
and cell motility. Thus, the balance of adhesive receptors ligated at
the cell surface during initial cell-matrix attachment serves to
regulate the type of substratum adhesion contact assembled and
subsequent cytoskeletal organization. A role for fascin microspikes in
cell motile behavior is indicated.
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