Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1986 Jan 1;102(1):160–178. doi: 10.1083/jcb.102.1.160

Cell adhesion and migration in the early vertebrate embryo: location and possible role of the putative fibronectin receptor complex

PMCID: PMC2114053  PMID: 2934401

Abstract

Using a combined in vivo and in vitro approach, we have analyzed the immunofluorescent localization and function of a 140,000-mol-wt glycoprotein complex implicated in cell adhesion to fibronectin (FN), with particular emphasis on neural crest cell adhesion and migration. This putative fibronectin receptor complex (FN-receptor) was detectable in almost all tissues derived from each of the three primary germ layers. It was present in both mesenchymal and epithelial cells, and was particularly enriched at sites close to concentrations of FN, e.g., at the basal surfaces of epithelial cells. It was also present on neural crest cells. The distribution and function of this putative receptor was then analyzed on individual cells in vitro. It was diffusely organized on highly locomotory neural crest cells and somitic fibroblasts. Both motile cell types also displayed relatively low numbers of focal contacts and microfilament bundles and limited amounts of localized vinculin, alpha-actinin, and endogenous FN. In contrast, the FN-receptor in stationary embryonic cells, i.e., somitic cells after long-term culture or ectodermal cells, existed in characteristic linear patterns generally co-distributed with alpha-actinin and fibers of endogenous FN. Anti-FN-receptor antibodies inhibited the adhesion to FN of motile embryonic cells, but not of stationary fibroblasts. However, these same antibodies adsorbed to substrata readily mediated adhesion and spreading of cells, but were much less effective for cell migration. Our results demonstrate a widespread occurrence in vivo of the putative FN-receptor, with high concentrations near FN. Embryonic cell migration was associated with a diffuse organization of this putative receptor on the cell surface in presumably labile adhesions, whereas stationary cells were anchored to the substratum at specific sites linked to the cytoskeleton near local concentrations of FN- receptor.

Full Text

The Full Text of this article is available as a PDF (2.7 MB).

Selected References

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

  1. Akiyama S. K., Yamada K. M. Synthetic peptides competitively inhibit both direct binding to fibroblasts and functional biological assays for the purified cell-binding domain of fibronectin. J Biol Chem. 1985 Sep 5;260(19):10402–10405. [PubMed] [Google Scholar]
  2. Akiyama S. K., Yamada K. M. The interaction of plasma fibronectin with fibroblastic cells in suspension. J Biol Chem. 1985 Apr 10;260(7):4492–4500. [PubMed] [Google Scholar]
  3. Ali I. U., Mautner V., Lanza R., Hynes R. O. Restoration of normal morphology, adhesion and cytoskeleton in transformed cells by addition of a transformation-sensitive surface protein. Cell. 1977 May;11(1):115–126. doi: 10.1016/0092-8674(77)90322-1. [DOI] [PubMed] [Google Scholar]
  4. Aplin J. D., Hughes R. C., Jaffe C. L., Sharon N. Reversible cross-linking of cellular components of adherent fibroblasts to fibronectin and lectin-coated substrata. Exp Cell Res. 1981 Aug;134(2):488–494. doi: 10.1016/0014-4827(81)90453-5. [DOI] [PubMed] [Google Scholar]
  5. Boucaut J. C., Darribere T. Fibronectin in early amphibian embryos. Migrating mesodermal cells contact fibronectin established prior to gastrulation. Cell Tissue Res. 1983;234(1):135–145. doi: 10.1007/BF00217407. [DOI] [PubMed] [Google Scholar]
  6. Boucaut J. C., Darribère T., Boulekbache H., Thiery J. P. Prevention of gastrulation but not neurulation by antibodies to fibronectin in amphibian embryos. 1984 Jan 26-Feb 1Nature. 307(5949):364–367. doi: 10.1038/307364a0. [DOI] [PubMed] [Google Scholar]
  7. Boucaut J. C., Darribère T., Poole T. J., Aoyama H., Yamada K. M., Thiery J. P. Biologically active synthetic peptides as probes of embryonic development: a competitive peptide inhibitor of fibronectin function inhibits gastrulation in amphibian embryos and neural crest cell migration in avian embryos. J Cell Biol. 1984 Nov;99(5):1822–1830. doi: 10.1083/jcb.99.5.1822. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Brackenbury R., Thiery J. P., Rutishauser U., Edelman G. M. Adhesion among neural cells of the chick embryo. I. An immunological assay for molecules involved in cell-cell binding. J Biol Chem. 1977 Oct 10;252(19):6835–6840. [PubMed] [Google Scholar]
  9. Bronner-Fraser M. Alterations in neural crest migration by a monoclonal antibody that affects cell adhesion. J Cell Biol. 1985 Aug;101(2):610–617. doi: 10.1083/jcb.101.2.610. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Brown P. J., Juliano R. L. Selective inhibition of fibronectin-mediated cell adhesion by monoclonal antibodies to a cell-surface glycoprotein. Science. 1985 Jun 21;228(4706):1448–1451. doi: 10.1126/science.4012302. [DOI] [PubMed] [Google Scholar]
  11. Burridge K., Feramisco J. R. Microinjection and localization of a 130K protein in living fibroblasts: a relationship to actin and fibronectin. Cell. 1980 Mar;19(3):587–595. doi: 10.1016/s0092-8674(80)80035-3. [DOI] [PubMed] [Google Scholar]
  12. Chapman A. E. Characterization of a 140Kd cell surface glycoprotein involved in myoblast adhesion. J Cell Biochem. 1984;25(2):109–121. doi: 10.1002/jcb.240250206. [DOI] [PubMed] [Google Scholar]
  13. Chen W. T., Greve J. M., Gottlieb D. I., Singer S. J. Immunocytochemical localization of 140 kD cell adhesion molecules in cultured chicken fibroblasts, and in chicken smooth muscle and intestinal epithelial tissues. J Histochem Cytochem. 1985 Jun;33(6):576–586. doi: 10.1177/33.6.3889142. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Chen W. T. Mechanism of retraction of the trailing edge during fibroblast movement. J Cell Biol. 1981 Jul;90(1):187–200. doi: 10.1083/jcb.90.1.187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Chen W. T., Singer S. J. Fibronectin is not present in the focal adhesions formed between normal cultured fibroblasts and their substrata. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7318–7322. doi: 10.1073/pnas.77.12.7318. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Couchman J. R., Rees D. A., Green M. R., Smith C. G. Fibronectin has a dual role in locomotion and anchorage of primary chick fibroblasts and can promote entry into the division cycle. J Cell Biol. 1982 May;93(2):402–410. doi: 10.1083/jcb.93.2.402. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. Critchley D. R., England M. A., Wakely J., Hynes R. O. Distribution of fibronectin in the ectoderm of gastrulating chick embryos. Nature. 1979 Aug 9;280(5722):498–500. doi: 10.1038/280498a0. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Decker C., Greggs R., Duggan K., Stubbs J., Horwitz A. Adhesive multiplicity in the interaction of embryonic fibroblasts and myoblasts with extracellular matrices. J Cell Biol. 1984 Oct;99(4 Pt 1):1398–1404. doi: 10.1083/jcb.99.4.1398. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Duband J. L., Thiery J. P. Appearance and distribution of fibronectin during chick embryo gastrulation and neurulation. Dev Biol. 1982 Dec;94(2):337–350. doi: 10.1016/0012-1606(82)90352-9. [DOI] [PubMed] [Google Scholar]
  24. Duband J. L., Thiery J. P. Distribution of fibronectin in the early phase of avian cephalic neural crest cell migration. Dev Biol. 1982 Oct;93(2):308–323. doi: 10.1016/0012-1606(82)90120-8. [DOI] [PubMed] [Google Scholar]
  25. Geiger B., Tokuyasu K. T., Dutton A. H., Singer S. J. Vinculin, an intracellular protein localized at specialized sites where microfilament bundles terminate at cell membranes. Proc Natl Acad Sci U S A. 1980 Jul;77(7):4127–4131. doi: 10.1073/pnas.77.7.4127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Greve J. M., Gottlieb D. I. Monoclonal antibodies which alter the morphology of cultured chick myogenic cells. J Cell Biochem. 1982;18(2):221–229. doi: 10.1002/jcb.1982.240180209. [DOI] [PubMed] [Google Scholar]
  27. Hasegawa T., Hasegawa E., Chen W. T., Yamada K. M. Characterization of a membrane-associated glycoprotein complex implicated in cell adhesion to fibronectin. J Cell Biochem. 1985;28(4):307–318. doi: 10.1002/jcb.240280409. [DOI] [PubMed] [Google Scholar]
  28. Heasman J., Hynes R. O., Swan A. P., Thomas V., Wylie C. C. Primordial germ cells of Xenopus embryos: the role of fibronectin in their adhesion during migration. Cell. 1981 Dec;27(3 Pt 2):437–447. doi: 10.1016/0092-8674(81)90385-8. [DOI] [PubMed] [Google Scholar]
  29. Heggeness M. H., Ash J. F., Singer S. J. Transmembrane linkage of fibronectin to intracellular actin-containing filaments in cultured human fibroblasts. Ann N Y Acad Sci. 1978 Jun 20;312:414–417. doi: 10.1111/j.1749-6632.1978.tb16822.x. [DOI] [PubMed] [Google Scholar]
  30. Horwitz A., Duggan K., Greggs R., Decker C., Buck C. The cell substrate attachment (CSAT) antigen has properties of a receptor for laminin and fibronectin. J Cell Biol. 1985 Dec;101(6):2134–2144. doi: 10.1083/jcb.101.6.2134. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. 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]
  32. Hynes R. O., Yamada K. M. Fibronectins: multifunctional modular glycoproteins. J Cell Biol. 1982 Nov;95(2 Pt 1):369–377. doi: 10.1083/jcb.95.2.369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Kleinman H. K., Martin G. R., Fishman P. H. Ganglioside inhibition of fibronectin-mediated cell adhesion to collagen. Proc Natl Acad Sci U S A. 1979 Jul;76(7):3367–3371. doi: 10.1073/pnas.76.7.3367. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Knudsen K. A., Horwitz A. F., Buck C. A. A monoclonal antibody identifies a glycoprotein complex involved in cell-substratum adhesion. Exp Cell Res. 1985 Mar;157(1):218–226. doi: 10.1016/0014-4827(85)90164-8. [DOI] [PubMed] [Google Scholar]
  35. Knudsen K. A., Rao P. E., Damsky C. H., Buck C. A. Membrane glycoproteins involved in cell--substratum adhesion. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6071–6075. doi: 10.1073/pnas.78.10.6071. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. 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]
  37. Laterra J., Silbert J. E., Culp L. A. Cell surface heparan sulfate mediates some adhesive responses to glycosaminoglycan-binding matrices, including fibronectin. J Cell Biol. 1983 Jan;96(1):112–123. doi: 10.1083/jcb.96.1.112. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Lohmann S. M., Walter U., Miller P. E., Greengard P., De Camilli P. Immunohistochemical localization of cyclic GMP-dependent protein kinase in mammalian brain. Proc Natl Acad Sci U S A. 1981 Jan;78(1):653–657. doi: 10.1073/pnas.78.1.653. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Mayer B. W., Jr, Hay E. D., Hynes R. O. Immunocytochemical localization of fibronectin in embryonic chick trunk and area vasculosa. Dev Biol. 1981 Mar;82(2):267–286. doi: 10.1016/0012-1606(81)90451-6. [DOI] [PubMed] [Google Scholar]
  40. Neff N. T., Lowrey C., Decker C., Tovar A., Damsky C., Buck C., Horwitz A. F. A monoclonal antibody detaches embryonic skeletal muscle from extracellular matrices. J Cell Biol. 1982 Nov;95(2 Pt 1):654–666. doi: 10.1083/jcb.95.2.654. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Newgreen D. F., Gibbins I. L., Sauter J., Wallenfels B., Wütz R. Ultrastructural and tissue-culture studies on the role of fibronectin, collagen and glycosaminoglycans in the migration of neural crest cells in the fowl embryo. Cell Tissue Res. 1982;221(3):521–549. doi: 10.1007/BF00215700. [DOI] [PubMed] [Google Scholar]
  42. 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]
  43. Neyfakh A. A., Jr, Svitkina T. M. Isolation of focal contact membrane using saponin. Exp Cell Res. 1983 Dec;149(2):582–586. doi: 10.1016/0014-4827(83)90369-5. [DOI] [PubMed] [Google Scholar]
  44. Patel V. P., Ciechanover A., Platt O., Lodish H. F. Mammalian reticulocytes lose adhesion to fibronectin during maturation to erythrocytes. Proc Natl Acad Sci U S A. 1985 Jan;82(2):440–444. doi: 10.1073/pnas.82.2.440. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Patel V. P., Lodish H. F. Loss of adhesion of murine erythroleukemia cells to fibronectin during erythroid differentiation. Science. 1984 Jun 1;224(4652):996–998. doi: 10.1126/science.6585955. [DOI] [PubMed] [Google Scholar]
  46. 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]
  47. Rovasio R. A., Delouvee A., Yamada K. M., Timpl R., Thiery J. P. Neural crest cell migration: requirements for exogenous fibronectin and high cell density. J Cell Biol. 1983 Feb;96(2):462–473. doi: 10.1083/jcb.96.2.462. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Ruoslahti E., Engvall E., Hayman E. G. Fibronectin: current concepts of its structure and functions. Coll Relat Res. 1981;1(1):95–128. doi: 10.1016/s0174-173x(80)80011-2. [DOI] [PubMed] [Google Scholar]
  49. Sanders E. J. Ultrastructural immunocytochemical localization of fibronectin in the early chick embryo. J Embryol Exp Morphol. 1982 Oct;71:155–170. [PubMed] [Google Scholar]
  50. Sieber-Blum M., Sieber F., Yamada K. M. Cellular fibronectin promotes adrenergic differentiation of quail neural crest cells in vitro. Exp Cell Res. 1981 Jun;133(2):285–295. doi: 10.1016/0014-4827(81)90320-7. [DOI] [PubMed] [Google Scholar]
  51. Singer I. I. Association of fibronectin and vinculin with focal contacts and stress fibers in stationary hamster fibroblasts. J Cell Biol. 1982 Feb;92(2):398–408. doi: 10.1083/jcb.92.2.398. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. 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]
  53. Singer I. I. The fibronexus: a transmembrane association of fibronectin-containing fibers and bundles of 5 nm microfilaments in hamster and human fibroblasts. Cell. 1979 Mar;16(3):675–685. doi: 10.1016/0092-8674(79)90040-0. [DOI] [PubMed] [Google Scholar]
  54. Spiegel S., Yamada K. M., Hom B. E., Moss J., Fishman P. H. Fluorescent gangliosides as probes for the retention and organization of fibronectin by ganglioside-deficient mouse cells. J Cell Biol. 1985 Mar;100(3):721–726. doi: 10.1083/jcb.100.3.721. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Tarone G., Galetto G., Prat M., Comoglio P. M. Cell surface molecules and fibronectin-mediated cell adhesion: effect of proteolytic digestion of membrane proteins. J Cell Biol. 1982 Jul;94(1):179–186. doi: 10.1083/jcb.94.1.179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Thiery J. P., Duband J. L., Delouvée A. Pathways and mechanisms of avian trunk neural crest cell migration and localization. Dev Biol. 1982 Oct;93(2):324–343. doi: 10.1016/0012-1606(82)90121-x. [DOI] [PubMed] [Google Scholar]
  57. Thiery J. P. Mechanisms of cell migration in the vertebrate embryo. Cell Differ. 1984 Nov;15(1):1–15. doi: 10.1016/0045-6039(84)90024-1. [DOI] [PubMed] [Google Scholar]
  58. Tucker R. P., Edwards B. F., Erickson C. A. Tension in the culture dish: microfilament organization and migratory behavior of quail neural crest cells. Cell Motil. 1985;5(3):225–237. doi: 10.1002/cm.970050305. [DOI] [PubMed] [Google Scholar]
  59. Vincent M., Thiery J. P. A cell surface marker for neural crest and placodal cells: further evolution in peripheral and central nervous system. Dev Biol. 1984 Jun;103(2):468–481. doi: 10.1016/0012-1606(84)90334-8. [DOI] [PubMed] [Google Scholar]
  60. Wehland J., Osborn M., Weber K. Cell-to-substratum contacts in living cells: a direct correlation between interference-reflexion and indirect-immunofluorescence microscopy using antibodies against actin and alpha-actinin. J Cell Sci. 1979 Jun;37:257–273. doi: 10.1242/jcs.37.1.257. [DOI] [PubMed] [Google Scholar]
  61. Willingham M. C., Yamada K. M., Yamada S. S., Pouysségur J., Pastan I. Microfilament bundles and cell shape are related to adhesiveness to substratum and are dissociable from growth control in cultured fibroblasts. Cell. 1977 Mar;10(3):375–380. doi: 10.1016/0092-8674(77)90024-1. [DOI] [PubMed] [Google Scholar]
  62. Wylie C. C., Heasman J. Effects of the substratum on the migration of primordial germ cells. Philos Trans R Soc Lond B Biol Sci. 1982 Nov 4;299(1095):177–183. doi: 10.1098/rstb.1982.0124. [DOI] [PubMed] [Google Scholar]
  63. Yamada K. M., Kennedy D. W. Dualistic nature of adhesive protein function: fibronectin and its biologically active peptide fragments can autoinhibit fibronectin function. J Cell Biol. 1984 Jul;99(1 Pt 1):29–36. doi: 10.1083/jcb.99.1.29. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES