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Modulation of expression and assembly of vinculin during in vitro fibrillar collagen-induced angiogenesis and its reversal.
Deroanne, Christophe; Colige, Alain; Nusgens, Betty et al.
1996In Experimental Cell Research, 224 (2), p. 215-23
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Keywords :
Actins/metabolism; Blotting, Northern; Blotting, Western; Cell Adhesion/genetics; Cell Adhesion Molecules/drug effects/metabolism; Cell Differentiation/drug effects/physiology; Collagen/pharmacology; Cytoskeleton/drug effects/metabolism; Endothelium/blood supply/metabolism; Epidermis/cytology; Fibroblasts/metabolism; Gene Expression/drug effects/physiology; Humans; Neovascularization, Physiologic/drug effects/genetics; Polymers/pharmacology; RNA, Messenger/metabolism; Umbilical Veins/cytology; Vinculin/genetics/metabolism
Abstract :
[en] A model of collagen-induced in vitro angiogenesis was used to investigate the modulation of expression and assembly of focal adhesion plaque-associated proteins during the process of differentiation. Human umbilical vein endothelial cells (HUVEC), first attached on an adhesive substratum (gelatin-, fibronectin-, or laminin-coated dish) or adherent collagen gel and then covered by an overlaying collagen get, organized within 3-4 days in tube-like structures (TLS). Removing the overlaying collagen gel from fully differentiated HUVEC induced a reversion of the process and HUVEC returned to a monolayer pattern. Modulations of focal adhesion-associated proteins occurring in HUVEC during the in vitro differentiation process and its reversal were investigated by Western blot analysis. A significant decrease of expression of vinculin, the integrin alpha2 subunit, talin, alpha-actinin, and actin was observed in TLS whereas the amount of FVIII-related antigen did not vary as compared to control monolayer cultures. During reversal, all the reduced proteins were markedly reexpressed. Human skin fibroblasts (HSF), submitted to the same experimental conditions, did not form TLS. Most of the focal adhesion proteins in HSF were similarly modulated by an overlaying collagen gel with the exception of vinculin, which was not modified. This particular protein was therefore more thoroughly investigated. In a nondifferentiated monolayer of HUVEC, a significant proportion of vinculin was organized into a detergent-resistant juxtamembranous structure (focal adhesion plaque) which disassembled early in TLS formation and reassembled during the reversal of the process. The reduction of vinculin during TLS formation was preceded by a downregulation of its mRNA while this mRNA was upregulated during reversal of the morphotype. These results suggest that the modulations of the cytoskeletal and focal adhesion proteins and more specifically of vinculin coupled to its subcellular redistribution are critical and early events in the cascade of mechanochemical signaling during in vitro angiogenesis induced by fibrillar collagen.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Deroanne, Christophe ;  Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Laboratoire des tissus conjonctifs
Colige, Alain ;  Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Laboratoire des tissus conjonctifs
Nusgens, Betty ;  Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Laboratoire de Biologie des Tissus Conjonctifs
Lapiere, C. M.;  Université de Liège - ULiège
Language :
English
Title :
Modulation of expression and assembly of vinculin during in vitro fibrillar collagen-induced angiogenesis and its reversal.
Publication date :
1996
Journal title :
Experimental Cell Research
ISSN :
0014-4827
eISSN :
1090-2422
Publisher :
Academic Press, Orlando, United States - Florida
Volume :
224
Issue :
2
Pages :
215-23
Peer reviewed :
Peer Reviewed verified by ORBi
Available on ORBi :
since 18 June 2010

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