The Journal of Cell Biology, Vol 109, 1795-1805, Copyright © 1989 by The Rockefeller University Press

ARTICLES

Cell adhesion to fibronectin and tenascin: quantitative measurements of initial binding and subsequent strengthening response

MM Lotz, CA Burdsal, HP Erickson and DR McClay
Department of Zoology, Duke University, Durham, North Carolina 27706.

Cell-substratum adhesion strengths have been quantified using fibroblasts and glioma cells binding to two extracellular matrix proteins, fibronectin and tenascin. A centrifugal force-based adhesion assay was used for the adhesive strength measurements, and the corresponding morphology of the adhesions was visualized by interference reflection microscopy. The initial adhesions as measured at 4 degrees C were on the order of 10(-5)dynes/cell and did not involve the cytoskeleton. Adhesion to fibronectin after 15 min at 37 degrees C were more than an order of magnitude stronger; the strengthening response required cytoskeletal involvement. By contrast to the marked strengthening of adhesion to FN, adhesion to TN was unchanged or weakened after 15 min at 37 degrees C. The absolute strength of adhesion achieved varied according to protein and cell type. When a mixed substratum of fibronectin and tenascin was tested, the presence of tenascin was found to reduce the level of the strengthening of cell adhesion normally observed at 37 degrees C on a substratum of fibronectin alone. Parallel analysis of corresponding interference reflection micrographs showed that differences in the area of cell surface within 10-15 nm of the substratum correlated closely with each of the changes in adhesion observed: after incubation for 15 min on fibronectin at 37 degrees C, glioma cells increased their surface area within close contact to the substrate by integral to 125- fold. Cells on tenascin did not increase their surface area of contact. The increased surface area of contact and the inhibitory activity of cytochalasin b suggest that the adhesive "strengthening" in the 15 min after initial binding brings additional adhesion molecules into the adhesive site and couples the actin cytoskeleton to the adhesion complex.
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• Sagvolden, G., Giaever, I., Pettersen, E. O., Feder, J. (1999). Cell adhesion force microscopy. Proc. Natl. Acad. Sci. USA 96: 471-476 [Abstract] [Full Text]  
• Mackie, E., Tucker, R. (1999). The tenascin-C knockout revisited. J. Cell Sci. 112: 3847-3853 [Abstract]  
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• LaFleur, D. W., Chiang, J., Fagin, J. A., Schwartz, S. M., Shah, P. K., Wallner, K., Forrester, J. S., Sharifi, B. G. (1997). Aortic Smooth Muscle Cells Interact with Tenascin-C through Its Fibrinogen-like Domain. J. Biol. Chem. 272: 32798-32803 [Abstract] [Full Text]  
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• Gagnoux-Palacios, L., Vailly, J., Durand-Clement, M., Wagner, E., Ortonne, J.-P., Meneguzzi, G. (1996). Functional Re-expression of Laminin-5 in Laminin-gamma 2-deficient Human Keratinocytes Modifies Cell Morphology, Motility, and Adhesion. J. Biol. Chem. 271: 18437-18444 [Abstract] [Full Text]  
• Murphy-Ullrich, J., Pallero, M., Boerth, N, Greenwood, J., Lincoln, T., Cornwell, T. (1996). Cyclic GMP-dependent protein kinase is required for thrombospondin and tenascin mediated focal adhesion disassembly. J. Cell Sci. 109: 2499-2508 [Abstract]  
• Prakobphol, A., Burdsal, C.A., Fisher, S.J. (1995). Quantifying the Strength of Bacterial Adhesive Interactions with Salivary Glycoproteins. JDR 74: 1212-1218 [Abstract]  
• Fischer, D., Chiquet-Ehrismann, R., Bernasconi, C., Chiquet, M. (1995). A Single Heparin Binding Region within the Fibrinogen-like Domain Is Functional in Chick Tenascin-C. J. Biol. Chem. 270: 3378-3384 [Abstract] [Full Text]  
• Adams, J. (1995). Formation of stable microspikes containing actin and the 55 kDa actin bundling protein, fascin, is a consequence of cell adhesion to thrombospondin-1: implications for the anti-adhesive activities of thrombospondin-1. J. Cell Sci. 108: 1977-1990 [Abstract]  
• Redick, S., Schwarzbauer, J. (1995). Rapid intracellular assembly of tenascin hexabrachions suggests a novel cotranslational process. J. Cell Sci. 108: 1761-1769 [Abstract]  
• Toyoizumi, R, Takeuchi, S (1995). The behavior of chick gastrula mesodermal cells under the unidirectional tractive force parallel to the substrata. J. Cell Sci. 108: 557-567 [Abstract]  
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• Tucker, R., Spring, J, Baumgartner, S, Martin, D, Hagios, C, Poss, P., Chiquet-Ehrismann, R (1994). Novel tenascin variants with a distinctive pattern of expression in the avian embryo. Development 120: 637-647 [Abstract]  
• Wehrle-Haller, B, Chiquet, M (1993). Dual function of tenascin: simultaneous promotion of neurite growth and inhibition of glial migration. J. Cell Sci. 106: 597-610 [Abstract]  
• Joshi, P, Chung, C., Aukhil, I, Erickson, H. (1993). Endothelial cells adhere to the RGD domain and the fibrinogen-like terminal knob of tenascin. J. Cell Sci. 106: 389-400 [Abstract]  
• Ingber, D. (1993). Cellular tensegrity: defining new rules of biological design that govern the cytoskeleton. J. Cell Sci. 104: 613-627  
• Tucker, R., Hammarback, J., Jenrath, D., Mackie, E., Xu, Y (1993). Tenascin expression in the mouse: in situ localization and induction in vitro by bFGF. J. Cell Sci. 104: 69-76 [Abstract]  
• Tucker, R. P. (1993). The in situ localization of tenascin splice variants and thrombospondin 2 mRNA in the avian embryo. Development 117: 347-358 [Abstract]  
• Sims, J. R., Karp, S., Ingber, D. E. (1992). Altering the cellular mechanical force balance results in integrated changes in cell, cytoskeletal and nuclear shape. J. Cell Sci. 103: 1215-1222 [Abstract]  
• Mackie, E. J., Tucker, R. P. (1992). Tenascin in bone morphogenesis: expression by osteoblasts and cell type-specific expression of splice variants. J. Cell Sci. 103: 765-771 [Abstract]  
• Boettiger, D., Lynch, L., Blystone, S., Huber, F. (2001). Distinct Ligand-binding Modes for Integrin alpha vbeta 3-Mediated Adhesion to Fibronectin versus Vitronectin. J. Biol. Chem. 276: 31684-31690 [Abstract] [Full Text]  

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