Reference : In vitro tubulogenesis of endothelial cells by relaxation of the coupling extracellular ...
Scientific journals : Article
Life sciences : Biochemistry, biophysics & molecular biology
http://hdl.handle.net/2268/63540
In vitro tubulogenesis of endothelial cells by relaxation of the coupling extracellular matrix-cytoskeleton.
English
Deroanne, Christophe mailto [Université de Liège - ULg > Département des sciences biomédicales et précliniques > Laboratoire des tissus conjonctifs >]
Lapiere, C. M. [Université de Liège - ULG > > > > Laboratoire de Biologie des Tissus Conjonctifs > >]
Nusgens, Betty mailto [Université de Liège - ULg > Département des sciences biomédicales et précliniques > Laboratoire de Biologie des Tissus Conjonctifs > >]
2001
Cardiovascular Research
Elsevier Science
49
3
647-58
Yes (verified by ORBi)
International
0008-6363
Amsterdam
The Netherlands
[en] Blotting, Western/methods ; Capillaries ; Cell Adhesion/physiology ; Cells, Cultured ; Collagen ; Cytoskeleton/physiology ; Drug Combinations ; Endothelium, Vascular/physiology/ultrastructure ; Extracellular Matrix/physiology ; Fibroblasts ; Humans ; Laminin ; Microscopy, Phase-Contrast ; Neovascularization, Physiologic/physiology ; Proteoglycans ; Stress, Mechanical ; Umbilical Veins
[en] OBJECTIVE: This investigation aimed at determining the importance of the rigidity of the adhesive support and the participation of the cytoskeleton in tubulogenesis of endothelial cells in vitro. METHODS: The morphotype, biosynthetic phenotype and cytoskeleton organization of human umbilical vein endothelial cells (HUVEC) were analyzed on supports of variable mechanical resistance. RESULTS: Western blot analysis revealed a strong reduction of the expression of actin and focal-adhesion plaque (FAP) proteins in HUVEC organized in tube-like structures (TLS) on soft matrigel or on matrigel co-polymerized with heat-denatured collagen as compared to HUVEC remaining in a monolayer pattern on rigid matrigel-coat or on matrigel co-polymerized with type I collagen. Human skin fibroblasts morphotype was not altered in these culture conditions and the pattern of FAP proteins and actin was not modulated. By using polyacrylamide gels polymerized with various concentrations of bis-acrylamide to modulate the mechanical resistance of the support and cross-linked to a constant amount of gelatin to provide an equal density of attachment sites, it was shown that the less rigid the support, the more endothelial cells switched to a tube-like pattern. Collagen type I-induced tubulogenesis was accompanied by a profound and reversible remodeling of the actin-FAP complex suggesting a weakening of the bridging between extracellular matrix (ECM) and the cytoskeleton. Human skin fibroblasts and smooth muscle cells, used as control cells, adhered strongly to the collagen, did not form TLS and their network of actin stress fibers was not remodeled. The inhibition of collagen type I-induced tubulogenesis by agents altering the actin cytoskeleton-FAP complex including calpain type I inhibitor, orthovanadate, KT5720 and jasplakinolide, further supports the determinant role of mechanical coupling between the cells and the matrix in tubulogenesis. CONCLUSIONS: A reduced tension between the endothelial cells and the extracellular matrix, originating in the support or within the cells is sufficient to trigger an intracellular signaling cascade leading to tubulogenesis, an event mimicking one of the last steps of angiogenesis.
Researchers
http://hdl.handle.net/2268/63540
http://cardiovascres.oxfordjournals.org/content/49/3/647.full.pdf+html
This is a pre-copy-editing, author-produced PDF of an article accepted for publication in Cardiovascular Research following peer review. The definitive publisher-authenticated version [Deroanne, C, Lapière, Ch. M., and Nusgens, B. In vitro tubulogenesis of endothelial cells by relaxation of the coupling extracellular matrix-cytoskeleton, Cardiovascular Res, 49(3): 647-658, 2001] is available online at http://cardiovascres.oxfordjournals.org/content/49/3/647.full.pdf+html

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