scholarly journals Revealing Early Steps of α2β1 Integrin-mediated Adhesion to Collagen Type I by Using Single-Cell Force Spectroscopy

2007 ◽  
Vol 18 (5) ◽  
pp. 1634-1644 ◽  
Author(s):  
Anna Taubenberger ◽  
David A. Cisneros ◽  
Jens Friedrichs ◽  
Pierre-Henri Puech ◽  
Daniel J. Muller ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Single Cell ◽  
Collagen Type ◽  
Chinese Hamster ◽  
Force Spectroscopy ◽  
Collagen Type I ◽  
Time Range ◽  
Type I ◽  
Actomyosin Contractility ◽  
Cell Force

We have characterized early steps of α2β1 integrin-mediated cell adhesion to a collagen type I matrix by using single-cell force spectroscopy. In agreement with the role of α2β1 as a collagen type I receptor, α2β1-expressing Chinese hamster ovary (CHO)-A2 cells spread rapidly on the matrix, whereas α2β1-negative CHO wild-type cells adhered poorly. Probing CHO-A2 cell detachment forces over a contact time range of 600 s revealed a nonlinear adhesion response. During the first 60 s, cell adhesion increased slowly, and forces associated with the smallest rupture events were consistent with the breakage of individual integrin–collagen bonds. Above 60 s, a fraction of cells rapidly switched into an activated adhesion state marked by up to 10-fold increased detachment forces. Elevated overall cell adhesion coincided with a rise of the smallest rupture forces above the value required to break a single-integrin–collagen bond, suggesting a change from single to cooperative receptor binding. Transition into the activated adhesion mode and the increase of the smallest rupture forces were both blocked by inhibitors of actomyosin contractility. We therefore propose a two-step mechanism for the establishment of α2β1-mediated adhesion as weak initial, single-integrin–mediated binding events are superseded by strong adhesive interactions involving receptor cooperativity and actomyosin contractility.

scholarly journals Stimulation of beta1 integrins on fibroblasts induces PDGF independent tyrosine phosphorylation of PDGF beta-receptors.

1996 ◽  
Vol 132 (4) ◽  
pp. 741-752 ◽  
Author(s):  
C Sundberg ◽  
K Rubin
Keyword(s):  
Cell Adhesion ◽  
Tyrosine Phosphorylation ◽  
Collagen Type ◽  
Collagen Gel ◽  
Collagen Type I ◽  
Receptor Activation ◽  
Type I ◽  
Integrin Subunit ◽  
Beta Receptor ◽  
Beta Receptors

We report that integrin-mediated signaling induces a rapid and transient tyrosine phosphorylation of platelet-derived growth factor (PDGF) beta-receptors in human diploid foreskin AG 1518 fibroblasts. A transient tyrosine phosphorylation of PDGF beta-receptors was evident one and two hours after cells had been plated on collagen type I and fibronectin, as well as on immobilized anti-integrin subunit IgG, but not on poly-L-lysine. In contrast EGF or PDGF alpha-receptors were not phosphorylated on tyrosine residues under these conditions. Tyrosine phosphorylation of PDGF beta-receptors induced by plating on collagen type I was inhibited by cytochalasin D and herbimycin A, unaffected by cycloheximide and enhanced by orthovanadate. Furthermore, a transient phosphorylation of PDGF beta-receptors occurred when AG 518 fibroblasts were cultured in three-dimensional collagen lattices or exposed to external strain exerted through centrifugation. The latter effect was evident already after two minutes. Clustering of cell surface beta1 integrins led to PDGF beta-receptor phosphorylation both in suspended and firmly attached AG 1518 fibroblasts. Plating of cells on collagen type I, fibronectin, and anti-beta1-integrin IgG resulted in the formation of PDGF beta-receptor aggregates as detected by immunofluorescence. Suramin or anti-PDGF-BB IgG had no effect on the plating-induced tyrosine phosphorylation of PDGF beta-receptors. PDGF-B chain mRNA, or protein, were not detected in AG 1518 fibroblasts. Our data suggest that a ligand-independent PDGF beta-receptor activation during cell adhesion and early phases of cell spreading is involved in integrin-mediated signaling in fibroblasts, and constitutes parts of a mechanism for cells to respond during the dynamic phases of externally applied tension as well as fibroblast-mediated tension during cell adhesion and collagen gel contraction.

Single Cell and Spheroid Collagen Type I Invasion Assay

2013 ◽  
pp. 13-35 ◽  
Author(s):  
Olivier De Wever ◽  
An Hendrix ◽  
Astrid De Boeck ◽  
Frank Eertmans ◽  
Wendy Westbroek ◽  
...  
Keyword(s):  
Single Cell ◽  
Collagen Type ◽  
Collagen Type I ◽  
Type I ◽  
Invasion Assay

scholarly journals Quantifying the effect of electric current on cell adhesion studied by single-cell force spectroscopy

2016 ◽  
Vol 11 (1) ◽  
pp. 011004 ◽  
Author(s):  
Leena Jaatinen ◽  
Eleanore Young ◽  
Jari Hyttinen ◽  
János Vörös ◽  
Tomaso Zambelli ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Single Cell ◽  
Electric Current ◽  
Force Spectroscopy ◽  
Cell Force

Stimulated single-cell force spectroscopy to quantify cell adhesion receptor crosstalk

PROTEOMICS ◽  
2010 ◽  
Vol 10 (7) ◽  
pp. 1455-1462 ◽  
Author(s):  
Jens Friedrichs ◽  
Jonne Helenius ◽  
Daniel J. Müller
Keyword(s):  
Cell Adhesion ◽  
Single Cell ◽  
Force Spectroscopy ◽  
Adhesion Receptor ◽  
Receptor Crosstalk ◽  
Cell Force ◽  
Cell Adhesion Receptor

scholarly journals Quantitative investigation of calcimimetic R568 on beta cell adhesion and mechanics using AFM single-cell force spectroscopy

FEBS Letters ◽  
2014 ◽  
Vol 588 (7) ◽  
pp. 1178-1183 ◽  
Author(s):  
Eleftherios Siamantouras ◽  
Claire E. Hills ◽  
Mustafa Y.G. Younis ◽  
Paul E. Squires ◽  
Kuo-Kang Liu
Keyword(s):  
Cell Adhesion ◽  
Beta Cell ◽  
Single Cell ◽  
Force Spectroscopy ◽  
Quantitative Investigation ◽  
Cell Force

scholarly journals Cell Adhesion on Dynamic Supramolecular Surfaces Probed by Fluid Force Microscopy-Based Single-Cell Force Spectroscopy

ACS Nano ◽  
2017 ◽  
Vol 11 (4) ◽  
pp. 3867-3874 ◽  
Author(s):  
Shrikrishnan Sankaran ◽  
Leena Jaatinen ◽  
Jenny Brinkmann ◽  
Tomaso Zambelli ◽  
Janos Vörös ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Single Cell ◽  
Force Spectroscopy ◽  
Fluid Force ◽  
Force Microscopy ◽  
Cell Force

Release of an invasion promoter E-cadherin fragment by matrilysin and stromelysin-1

2001 ◽  
Vol 114 (1) ◽  
pp. 111-118 ◽  
Author(s):  
V. Noe ◽  
B. Fingleton ◽  
K. Jacobs ◽  
H.C. Crawford ◽  
S. Vermeulen ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Cell Surface ◽  
Collagen Type ◽  
Cell Aggregation ◽  
Collagen Type I ◽  
Type I ◽  
Tissue Inhibitor Of Metalloproteinases ◽  
Dependent Cell ◽  
Mcf 7 ◽  
E Cadherin

The function of many transmembrane molecules can be altered by cleavage and subsequent release of their ectodomains. We have investigated ectodomain cleavage of the cell-cell adhesion and signal-transducing molecule E-cadherin. The E-cadherin ectodomain is constitutively shed from the surface of MCF-7 and MDCKts.srcC12 cells in culture. Release of the 80 kDa soluble E-cadherin fragment is stimulated by phorbol-12-myristate-13-acetate and is inhibited by overexpression of the tissue inhibitor of metalloproteinases-2. The metalloproteinases matrilysin and stromelysin-1 both cleave E-cadherin at the cell surface and release sE-CAD into the medium. The soluble E-cadherin fragment thus released inhibits E-cadherin functions in a paracrine way, as indicated by induction of invasion into collagen type I and inhibition of E-cadherin-dependent cell aggregation. Our results, therefore, suggest a novel mechanism by which metalloproteinases can influence invasion.

scholarly journals Nanostructured TiC Layer is Highly Suitable Surface for Adhesion, Proliferation and Spreading of Cells

2020 ◽  
Vol 5 (2) ◽  
pp. 29 ◽  
Author(s):  
Mariangela Lopreiato ◽  
Alessia Mariano ◽  
Rossana Cocchiola ◽  
Giovanni Longo ◽  
Pietro Dalla Vedova ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Titanium Carbide ◽  
Collagen Type ◽  
Collagen Type I ◽  
Bone Morphogenetic Protein 2 ◽  
Type I ◽  
Good Adhesion ◽  
Functional Coating ◽  
Adhesion Properties ◽  
Coated Glass

Cell culture is usually performed in 2D polymer surfaces; however, several studies are conducted with the aim to screen functional coating molecules to find substrates more suitable for cell adhesion and proliferation. The aim of this manuscript is to compare the cell adhesion and cytoskeleton organization of different cell types on different surfaces. Human primary fibroblasts, chondrocytes and osteoblasts isolated from patients undergoing surgery were seeded on polystyrene, poly-d-lysine-coated glass and titanium carbide slides and left to grow for several days. Then their cytoskeleton was analyzed, both by staining cells with phalloidin, which highlights actin fibers, and using Atomic Force Microscopy. We also monitored the production of Fibroblast Growth Factor-2, Bone Morphogenetic Protein-2 and Osteocalcin, using ELISA, and we highlighted production of Collagen type I in fibroblasts and osteoblasts and Collagen type II in chondrocytes by immunofluorescences. Fibroblasts, chondrocytes and osteoblasts showed both an improved proliferative activity and a good adhesion ability when cultured on titanium carbide slides, compared to polystyrene and poly-d-lysine-coated glass. In conclusion, we propose titanium carbide as a suitable surface to cultivate cells such as fibroblasts, chondrocytes and osteoblasts, allowing the preservation of their differentiated state and good adhesion properties.

scholarly journals Interactions of the neural cell adhesion molecule and the myelin-associated glycoprotein with collagen type I: involvement in fibrillogenesis.

1992 ◽  
Vol 116 (4) ◽  
pp. 1063-1070 ◽  
Author(s):  
R Probstmeier ◽  
T Fahrig ◽  
E Spiess ◽  
M Schachner
Keyword(s):  
Extracellular Matrix ◽  
Cell Adhesion ◽  
Adhesion Molecules ◽  
Cell Adhesion Molecule ◽  
Neural Cell Adhesion Molecule ◽  
Collagen Type ◽  
Collagen Type I ◽  
Type I ◽  
Neural Cell Adhesion ◽  
Myelin Associated Glycoprotein

To gain insights into the functional role of the molecular association between neural adhesion molecules and extracellular matrix constituents, soluble forms of the myelin-associated glycoprotein (MAG) and the neural cell adhesion molecule (N-CAM), representing most of the extracellular domains of the molecules, were investigated in their ability to modify fibrillogenesis of collagen type I. MAG and N-CAM retarded the rate of fibril formation, as measured by changes in turbidity, and increased the diameter of the fibrils formed, but did not change the banding pattern when compared to collagen type I in the absence of adhesion molecules. Scatchard plot analysis of the binding of MAG and N-CAM to the fibril-forming collagen types I, II, III, and V suggest one binding site for N-CAM and two binding sites for MAG. Binding of MAG, but not of N-CAM, to collagen type I was decreased during fibril formation, probably due to a reduced accessibility of one binding site for MAG during fibrillogenesis. These results indicate that the neural adhesion molecules can influence the configuration of extracellular matrix constituents, thus, implicating them in the modulation of cell-substrate interactions.

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