Adhesion-induced intracellular signalling in endothelial cells depends on the nature of the matrix.
The adhesion of a human microvascular endothelial cell line to its own matrix was studied in comparison with adhesion of the same cells to fibronectin or thrombospondin-1. These endothelial cells adhered preferentially to their matrix whereas an equal cell number was attached to fibronectin or thrombospondin-1. The adhesion of cells to thrombospondin-1 was mediated by the N-terminal heparin binding domain of thrombospondin-1 as shown by the use of a recombinant fragment, N18. Cells adhering to their matrix displayed a morphology and a cytoskeleton organization very similar to that observed in vivo with an apical immunostaining for actin stress fibers and a fine basal labeling for vinculin. Cells on fibronectin were extensively spread and rapidly assembled stress fibers and focal contacts. Cells adherent to thrombospondin-1 presented large lamellae rich in actin but devoid of vinculin and only few actin fibers were observed. Depending on the substratum used, adhering endothelial cells displayed also different tyrosine phosphorylation patterns on electrophoresis. Our observations indicate that endothelial cells adhering to their matrix present an activation state intermediate between that induced by a "hyperadhesive" protein like fibronectin and that generated by a moderate, indeed anti-adhesive, protein like thrombospondin-1.
Berge
N
N
U353 INSERM: Protéines Adhésives et Protéases des Cellules Vasculaires et Sanguines, Hôpital Saint-Louis, Paris, France.
Loganadane
L D
LD
Vassy
J
J
Monnet
E
E
Legrand
C
C
Fauvel-Lafeve
F
F
D000199:Actins / Q000378:metabolism; D002448:Cell Adhesion; D002460:Cell Line; D004730:Endothelium, Vascular / Q000502:physiology*; D005109:Extracellular Matrix / Q000502:physiology*; D005353:Fibronectins / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D018613:Microscopy, Confocal; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D015398:Signal Transduction; D019700:Thrombospondin 1 / Q000378:metabolism; D013997:Time Factors; D014443:Tyrosine / Q000378:metabolism; D016596:Vinculin / Q000378:metabolism
D000199:Actins; D005353:Fibronectins; D011993:Recombinant Fusion Proteins; D019700:Thrombospondin 1; D016596:Vinculin; D019000:Phosphotyrosine; D014443:Tyrosine