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. 1986 Dec 1;103(6):2697–2706. doi: 10.1083/jcb.103.6.2697

Focal adhesion sites and the removal of substratum-bound fibronectin

PMCID: PMC2114597  PMID: 2947902

Abstract

Fibronectin was not removed from the substratum beneath focal adhesion sites when fibroblasts spread in serum-free medium on adsorbed fibronectin substrata, or when fibroblasts spread in serum-containing medium on covalently cross-linked fibronectin substrata. Under these conditions, there was colocalization between 140-kD fibronectin receptors and focal adhesion sites. It was concluded that removal of adsorbed fibronectin from beneath focal adhesion sites was a mechanical process that required serum. The effect of serum was nonspecific since serum could be replaced by equivalent concentrations of serum albumin, ovalbumin, or gamma globulins. Quantitative measurements indicated that the presence of proteins in the incubation medium weakens the interaction of fibronectin with the substratum, thereby allowing the adsorbed protein to be removed from the substratum at sites of high stress. After removing fibronectin from the substratum, cells reorganized this material into patches and fibrils beneath cells, and the reorganized fibronectin colocalized with fibronectin receptors. Some of the patches of fibronectin were phagocytosed. The fibronectin fibrils were observed to be in register with actin filament bundles and sometimes translocated to the upper cell surfaces. It is proposed that removal of fibronectin from beneath focal adhesion sites is an example of how cells can modify their extracellular matrices through contractile activity.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abercrombie M., Heaysman J. E., Pegrum S. M. The locomotion of fibroblasts in culture. IV. Electron microscopy of the leading lamella. Exp Cell Res. 1971 Aug;67(2):359–367. doi: 10.1016/0014-4827(71)90420-4. [DOI] [PubMed] [Google Scholar]
  2. Aplin J. D., Hughes R. C. Protein-derivatised glass coverslips for the study of cell-to substratum adhesion. Anal Biochem. 1981 May 1;113(1):144–148. doi: 10.1016/0003-2697(81)90057-9. [DOI] [PubMed] [Google Scholar]
  3. Avnur Z., Geiger B. The removal of extracellular fibronectin from areas of cell-substrate contact. Cell. 1981 Jul;25(1):121–132. doi: 10.1016/0092-8674(81)90236-1. [DOI] [PubMed] [Google Scholar]
  4. Badley R. A., Woods A., Smith C. G., Rees D. A. Actomyosin relationships with surface features in fibroblast adhesion. Exp Cell Res. 1980 Apr;126(2):263–272. doi: 10.1016/0014-4827(80)90264-5. [DOI] [PubMed] [Google Scholar]
  5. Bell E., Ivarsson B., Merrill C. Production of a tissue-like structure by contraction of collagen lattices by human fibroblasts of different proliferative potential in vitro. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1274–1278. doi: 10.1073/pnas.76.3.1274. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Birchmeier C., Kreis T. E., Eppenberger H. M., Winterhalter K. H., Birchmeier W. Corrugated attachment membrane in WI-38 fibroblasts: alternating fibronectin fibers and actin-containing focal contacts. Proc Natl Acad Sci U S A. 1980 Jul;77(7):4108–4112. doi: 10.1073/pnas.77.7.4108. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Brash J. L., Uniyal S., Samak Q. Exchange of albumin adsorbed on polymer surfaces. Trans Am Soc Artif Intern Organs. 1974;20A:69–76. [PubMed] [Google Scholar]
  8. Byers H. R., White G. E., Fujiwara K. Organization and function of stress fibers in cells in vitro and in situ. A review. Cell Muscle Motil. 1984;5:83–137. doi: 10.1007/978-1-4684-4592-3_2. [DOI] [PubMed] [Google Scholar]
  9. Chen W. T., Hasegawa E., Hasegawa T., Weinstock C., Yamada K. M. Development of cell surface linkage complexes in cultured fibroblasts. J Cell Biol. 1985 Apr;100(4):1103–1114. doi: 10.1083/jcb.100.4.1103. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Chen W. T., Singer S. J. Immunoelectron microscopic studies of the sites of cell-substratum and cell-cell contacts in cultured fibroblasts. J Cell Biol. 1982 Oct;95(1):205–222. doi: 10.1083/jcb.95.1.205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Chernousov M. A., Metsis M. L., Koteliansky V. E. Studies of extracellular fibronectin matrix formation with fluoresceinated fibronectin and fibronectin fragments. FEBS Lett. 1985 Apr 22;183(2):365–369. doi: 10.1016/0014-5793(85)80811-5. [DOI] [PubMed] [Google Scholar]
  12. Couchman J. R., Rees D. A. The behaviour of fibroblasts migrating from chick heart explants: changes in adhesion, locomotion and growth, and in the distribution of actomyosin and fibronectin. J Cell Sci. 1979 Oct;39:149–165. doi: 10.1242/jcs.39.1.149. [DOI] [PubMed] [Google Scholar]
  13. Damsky C. H., Knudsen K. A., Bradley D., Buck C. A., Horwitz A. F. Distribution of the cell substratum attachment (CSAT) antigen on myogenic and fibroblastic cells in culture. J Cell Biol. 1985 May;100(5):1528–1539. doi: 10.1083/jcb.100.5.1528. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Fox C. H., Cottler-Fox M. H., Yamada K. M. The distribution of fibronectin in attachment sites of chick fibroblasts. Exp Cell Res. 1980 Dec;130(2):477–481. doi: 10.1016/0014-4827(80)90031-2. [DOI] [PubMed] [Google Scholar]
  15. Geiger B. A 130K protein from chicken gizzard: its localization at the termini of microfilament bundles in cultured chicken cells. Cell. 1979 Sep;18(1):193–205. doi: 10.1016/0092-8674(79)90368-4. [DOI] [PubMed] [Google Scholar]
  16. Geiger B., Avnur Z., Kreis T. E., Schlessinger J. The dynamics of cytoskeletal organization in areas of cell contact. Cell Muscle Motil. 1984;5:195–234. doi: 10.1007/978-1-4684-4592-3_5. [DOI] [PubMed] [Google Scholar]
  17. Giancotti F. G., Comoglio P. M., Tarone G. A 135,000 molecular weight plasma membrane glycoprotein involved in fibronectin-mediated cell adhesion. Immunofluorescence localization in normal and RSV-transformed fibroblasts. Exp Cell Res. 1986 Mar;163(1):47–62. doi: 10.1016/0014-4827(86)90557-4. [DOI] [PubMed] [Google Scholar]
  18. Grinnell F. Cellular adhesiveness and extracellular substrata. Int Rev Cytol. 1978;53:65–144. doi: 10.1016/s0074-7696(08)62241-x. [DOI] [PubMed] [Google Scholar]
  19. Grinnell F., Feld M. K. Adsorption characteristics of plasma fibronectin in relationship to biological activity. J Biomed Mater Res. 1981 May;15(3):363–381. doi: 10.1002/jbm.820150308. [DOI] [PubMed] [Google Scholar]
  20. Grinnell F., Geiger B. Interaction of fibronectin-coated beads with attached and spread fibroblasts. Binding, phagocytosis, and cytoskeletal reorganization. Exp Cell Res. 1986 Feb;162(2):449–461. doi: 10.1016/0014-4827(86)90349-6. [DOI] [PubMed] [Google Scholar]
  21. Grinnell F., Lamke C. R. Reorganization of hydrated collagen lattices by human skin fibroblasts. J Cell Sci. 1984 Mar;66:51–63. doi: 10.1242/jcs.66.1.51. [DOI] [PubMed] [Google Scholar]
  22. Grinnell F. Visualization of cell-substratum adhesion plaques by antibody exclusion. Cell Biol Int Rep. 1980 Nov;4(11):1031–1036. doi: 10.1016/0309-1651(80)90176-9. [DOI] [PubMed] [Google Scholar]
  23. Haas R., Culp L. A. Properties and fate of plasma fibronectin bound to the tissue culture substratum. J Cell Physiol. 1982 Nov;113(2):289–297. doi: 10.1002/jcp.1041130217. [DOI] [PubMed] [Google Scholar]
  24. Hynes R. O., Destree A. T. Relationships between fibronectin (LETS protein) and actin. Cell. 1978 Nov;15(3):875–886. doi: 10.1016/0092-8674(78)90272-6. [DOI] [PubMed] [Google Scholar]
  25. Izzard C. S., Lochner L. R. Cell-to-substrate contacts in living fibroblasts: an interference reflexion study with an evaluation of the technique. J Cell Sci. 1976 Jun;21(1):129–159. doi: 10.1242/jcs.21.1.129. [DOI] [PubMed] [Google Scholar]
  26. Kolega J., Shure M. S., Chen W. T., Young N. D. Rapid cellular translocation is related to close contacts formed between various cultured cells and their substrata. J Cell Sci. 1982 Apr;54:23–34. doi: 10.1242/jcs.54.1.23. [DOI] [PubMed] [Google Scholar]
  27. Lark M. W., Laterra J., Culp L. A. Close and focal contact adhesions of fibroblasts to a fibronectin-containing matrix. Fed Proc. 1985 Feb;44(2):394–403. [PubMed] [Google Scholar]
  28. Mangeat P., Burridge K. Actin-membrane interaction in fibroblasts: what proteins are involved in this association? J Cell Biol. 1984 Jul;99(1 Pt 2):95s–103s. doi: 10.1083/jcb.99.1.95s. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. McAbee D. D., Grinnell F. Fibronectin-mediated binding and phagocytosis of polystyrene latex beads by baby hamster kidney cells. J Cell Biol. 1983 Nov;97(5 Pt 1):1515–1523. doi: 10.1083/jcb.97.5.1515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Newgreen D., Thiery J. P. Fibronectin in early avian embryos: synthesis and distribution along the migration pathways of neural crest cells. Cell Tissue Res. 1980;211(2):269–291. doi: 10.1007/BF00236449. [DOI] [PubMed] [Google Scholar]
  31. Norton E. K., Izzard C. S. Fibronectin promotes formation of the close cell-to-substrate contact in cultured cells. Exp Cell Res. 1982 Jun;139(2):463–467. doi: 10.1016/0014-4827(82)90282-8. [DOI] [PubMed] [Google Scholar]
  32. Pytela R., Pierschbacher M. D., Ruoslahti E. Identification and isolation of a 140 kd cell surface glycoprotein with properties expected of a fibronectin receptor. Cell. 1985 Jan;40(1):191–198. doi: 10.1016/0092-8674(85)90322-8. [DOI] [PubMed] [Google Scholar]
  33. Singer I. I., Paradiso P. R. A transmembrane relationship between fibronectin and vinculin (130 kd protein): serum modulation in normal and transformed hamster fibroblasts. Cell. 1981 May;24(2):481–492. doi: 10.1016/0092-8674(81)90339-1. [DOI] [PubMed] [Google Scholar]
  34. Steinberg B. M., Smith K., Colozzo M., Pollack R. Establishment and transformation diminish the ability of fibroblasts to contract a native collagen gel. J Cell Biol. 1980 Oct;87(1):304–308. doi: 10.1083/jcb.87.1.304. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Stopak D., Harris A. K. Connective tissue morphogenesis by fibroblast traction. I. Tissue culture observations. Dev Biol. 1982 Apr;90(2):383–398. doi: 10.1016/0012-1606(82)90388-8. [DOI] [PubMed] [Google Scholar]
  36. Woods A., Couchman J. R., Johansson S., Hök M. Adhesion and cytoskeletal organisation of fibroblasts in response to fibronectin fragments. EMBO J. 1986 Apr;5(4):665–670. doi: 10.1002/j.1460-2075.1986.tb04265.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

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