Gastrulation pp 185-198 | Cite as

The Expression of Fibronectins and Integrins During Mesodermal Induction and Gastrulation in Xenopus

  • Douglas W. DeSimone
  • Jim C. Smith
  • James E. Howard
  • David G. Ransom
  • Karen Symes
Part of the Bodega Marine Laboratory Marine Science Series book series (BMSS)


Fibronectins (FNs) and integrins are first expressed in Xenopus embryos during the mid to late blastula stages. FN is synthesized in both animal and vegetal halves of the embryo but becomes localized to the roof of the blastocoel during gastrulation. Integrins are expressed in all regions of the early embryo. Structural heterogeneity of FN isoforms during embryogenesis occurs by alternative splicing of a common FN transcript, whereas integrin diversity is generated by the expression of several distinct integrin αβ heterodimers. The timing of expression for these molecules suggests that they may play important roles in supporting and/or controlling morphogenetic events in the early embryo.

We have investigated the roles played by these proteins in supporting the gastrulation-like movements that occur in animal pole tissue in response to mesoderminducing factors. Xenopus animal pole ectoderm was isolated from stage 8 embryos and exposed to the XTC mesoderm inducing factor (XTC-MIF; a Xenopus homologue of activin A). Animal pole ectoderm treated with XTC-MIF, like stage 10 dorsal marginal zone, will adhere and spread on FN coated surfaces. Uninduced animal pole ectoderm adheres poorly and does not spread on FN. The ability to spread on FN in response to XTC-MIF is also retained by single cells derived from dissociated animal pole tissue. This defines one of the few mesoderm-specific responses to induction that has been demonstrated for single cells. FN-mediated cell spreading is inhibited in the presence of the synthetic peptide Gly-Arg-Gly-Asp-Ser-Pro (GRGDSP), which corresponds to one of the active cell binding sites on the FN molecule. However, the gastrulation-like movements associated with elongation of XTC-MIF induced animal pole ectoderm are not inhibited by the GRGDSP peptide. These results indicate that convergent extension does not depend on cell adhesion to FN. Furthermore, scanning electron microscopy and cell marking techniques suggest that although cellular activity is enhanced following induction, no long range cell mixing occurs during elongation of induced explants. We are now investigating whether the changes in cell adhesion observed following induction with XTC-MIF are controlled by the expression of integrin receptors and ECM molecules such as FN.


Mesodermal Cell Xenopus Embryo Animal Pole Chick Embryo Fibroblast Convergent Extension 


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  1. Adams, J.C and F.M. Watt. 1990. Changes in keratinocyte adhesion during terminal differentiation: Reduction in fibronectin binding precedes α5βi integrin loss from the cell surface. Cell 63:425–435.PubMedCrossRefGoogle Scholar
  2. Albelda, S.M. and C.A. Buck. 1990. Integrins and other cell adhesion molecules. FASEB J. 4:2868–2880.PubMedGoogle Scholar
  3. Boucaut, J.-C, T. Darribère, H. Boulekbache, and J.-P. Thiery. 1984a. Prevention of gastrulation but not neurulation by antibody to fibronectin in amphibian embryos. Nature 307:364–367.PubMedCrossRefGoogle Scholar
  4. Boucaut, J.-C, T. Darribère, T.J. Poole, H. Aoyama, K.M. Yamada, and J.-P. Thiery. 1984b. 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. 99:1822–1830.PubMedCrossRefGoogle Scholar
  5. Boucaut, J.-C, T. Darribère, D. Shi, J.-F. Riou, K.E. Johnson, and M. Delarue. 1991. Amphibian Gastrulation: The Molecular Bases of Mesodermal Cell Migration in Urodele Embryos, p. 169–184. In: Gastrulation: Movements, Patterns, and Molecules. R. Keller, W.H. Clark, Jr., F. Griffin (Eds.). Plenum Press, New York.Google Scholar
  6. Cooke, J. and J.C. Smith. 1989. Gastrulation and larval pattern in Xenopus after blastocoelic injection of a Xenopus inducing factor: Experiments testing models for the normal organization of mesoderm. Dev. Biol. 131:383–400.PubMedCrossRefGoogle Scholar
  7. Cooke, J., J.C. Smith, E.J. Smith, and M. Yaqoob. 1987. The organization of mesodermal pattern in Xenopus laevis:Experiments using a Xenopus mesoderm-inducing factor. Development 101:893–908.PubMedGoogle Scholar
  8. Darribère, T., K. Guida, H. Larjava, K.E. Johnson, KM. Yamada, J.-P. Thiery, and J.-C. Boucaut. 1990. In vivo analyses of integrin β1 subunit function in fibronectin matrix assembly. J. Cell Biol.110:1813–1823.PubMedCrossRefGoogle Scholar
  9. Darribère, T., K.M. Yamada, K.E. Johnson, and J.-C. Boucaut. 1988. The 140 kD fibronectin receptor complex is required for mesodermal cell adhesion during gastrulation in the amphibian Pleurodeles waltlii. Dev. Biol. 126:182–194.PubMedCrossRefGoogle Scholar
  10. DeSimone, D.W. and R.O. Hynes. 1988. Xenopus laevis integrins: Structural conservation and evolutionary divergence of integrin β subunits. J. Biol. Chem. 263:5333–5340.PubMedGoogle Scholar
  11. Gimlich, R.L. and J. Braun. 1985. Improved fluorescent compound for tracing cell lineage. Dev. Biol. 109:509–514.PubMedCrossRefGoogle Scholar
  12. Heino, J., R.A. Ignotz, M.E. Hemler, C. Crouse, J. Massague. 1989. Regulation of cell adhesion receptors by transforming growth factor-β. Concomitant regulation of integrins that share a common β1 subunit. J. Biol. Chem. 264:380–388.PubMedGoogle Scholar
  13. Heino, J. and J. Massague. 1989. Transforming growth factor β switches the pattern of integrins expressed in MG-63 human osteosarcoma cells and causes a selective loss of adhesion to laminin. J. Biol. Chem. 264:21806–21811.PubMedGoogle Scholar
  14. Hynes, R.O. 1990. Fibronectins. Springer Verlag, New York.CrossRefGoogle Scholar
  15. Johnson, K.E., J.C. Boucaut, and D.W. DeSimone. 1991. The role of the extracellular matrix in amphibian gastrulation. Curr. Top. Dev. Biol. In press.Google Scholar
  16. Keller, R.E. and J. Hardin. 1987. Cell behavior during active cell rearrangement: Evidence and speculations. J. Cell Set Suppl 8:369–393.Google Scholar
  17. Keller, R.E. and P. Tibbetts. 1989. Mediolateral cell intercalation in the dorsal axial mesoderm of Xenopus laevis. Dev. Biol. 131:539–549.PubMedCrossRefGoogle Scholar
  18. Keller, R., J. Shih, and P. Wilson. 1991. Cell Motility, Control and Function of Convergence and Extension During Gastrulation in Xenopus. p. 101–120. In: Gastrulation: Movements, Patterns, and Molecules. R. Keller, W.H. Clark, Jr., F. Griffm (Eds.). Plenum Press, New York.Google Scholar
  19. Komazaki, S. 1988. Factors related to the initiation of cell migration along the inner surface of the blastocoelic wall during amphibian gastrulation. Cell Differ. 24:25–32.PubMedCrossRefGoogle Scholar
  20. Krieg, P.A. and D.A. Melton. 1987. In vitro synthesis with SP6 RNA polymerase. Methods Enzymol. 155:397–415.PubMedCrossRefGoogle Scholar
  21. Lee, G., R.O. Hynes, and M. Kirshner. 1984. Temporal and spatial regulation of fibronectin in early Xenopus development. Cell 36:729–740.PubMedCrossRefGoogle Scholar
  22. Marcantonio, E.E. and R.O. Hynes. 1988. Antibodies to the conserved cytoplasmic domain of the integrin β1 subunit react with proteins in vertebrates, invertebrates, and fungi. J. Cell Biol. 106:1765–1772.PubMedCrossRefGoogle Scholar
  23. Massague, J. 1990. The transforming growth factor-β family. Annu. Rev. Cell Biol. 6:597–641.PubMedCrossRefGoogle Scholar
  24. Melton, D.A. and R. Cortese. 1979. Transcription of cloned tRNA genes and nuclear partitioning of a tRNA precursor. Cell 18:1165–1172.PubMedCrossRefGoogle Scholar
  25. Nakatsuji, N. 1986. Presumptive mesodermal cells from Xenopus laevis gastrulae attach and migrate on substrata coated with fibronectin or laminin. J. Cell Sci. 86:109–118.PubMedGoogle Scholar
  26. Nakatsuji, N. and K.E. Johnson. 1983. Comparative study of extracellular fibrils on the ectodermal layer in gastrulae of five amphibian species. J. Cell Sci. 59:61–70.PubMedGoogle Scholar
  27. New, H.V. and J.C. Smith. 1990. Inductive interactions in early amphibian development. Curr. Opin. Cell Biol. 2:969–974.PubMedCrossRefGoogle Scholar
  28. Newport, J., and M. Kirschner. 1982. A major developmental transition in early Xenopus embryos. 1. Characterization and timing of cellular changes at the midblastula stage. Cell 30:675–686.PubMedCrossRefGoogle Scholar
  29. Nieuwkoop, P.D. 1969. The formation of mesoderm in urodelean amphibians. I. Induction by the endoderm. Wilhelm Roux’Arch. Entwicklungsmech. Org. 162:341–347.CrossRefGoogle Scholar
  30. Nieuwkoop, P.D. and J. Faber. 1967. Normal table of Xenopus laevis (Daudin). 2nd edition. North-Holland, Amsterdam.Google Scholar
  31. Nieuwkoop, P.D. and S. Sudarwati. 1971. Mesoderm formation in the Anuran Xenopus laevis (Daudin). Wilhelm Roux’s Arch. Dev. Biol. 166:189–204.CrossRefGoogle Scholar
  32. Pierschbacher, M.D. and E. Ruoslahti. 1984. Cell attachment activity of fibronectin can be duplicated by small synthetic fragments of the molecule. Nature 309:30–33.PubMedCrossRefGoogle Scholar
  33. Pytela, R., M.D. Pierschbacher, and E. Ruoslahti. 1985. Identification and isolation of a 140 kd cell surface glycoprotein with properties expected of a fibronectin receptor. Cell 40:191–198.PubMedCrossRefGoogle Scholar
  34. Ransom, D.G. and D.W. DeSimone. 1990. Cloning and characterization of multiple integrin αand β subunits expressed in Xenopus embryos. J. Cell Biol. 111:142a.Google Scholar
  35. Shi, D.-L., T. Darribère, K.E. Johnson, and J.-C. Boucaut. 1989. Initiation of mesodermal cell migration and spreading relative to gastrulation in the urodele amphibian Pleurodeles waltl. Development 105:351–363.Google Scholar
  36. Slack, J.M. 1984. Regional biosynthetic markers in the early amphibian embryo. J. Embryol Exp. Morphol. 80:289–319.PubMedGoogle Scholar
  37. Smith, J.C., K. Symes, R.O. Hynes, and D.W. DeSimone. 1990. Mesoderm induction and the control of gastrulation in Xenopus laevis: The roles of fibronectin and integrins. Development 108:229–238.PubMedGoogle Scholar
  38. Smith, J.C., M. Yaqoob, and K. Symes. 1988. Purification, partial characterization and biological properties of the XTC mesoderm inducing factor. Development 103:591–600.PubMedGoogle Scholar
  39. Symes, K. and J.C. Smith. 1987. Gastrulation movements provide an early marker of mesoderm induction in Xenopus laevis. Development 101:339–349.Google Scholar
  40. Tickle, C. and J.P. Trinkaus. 1973. Change in surface extensibility of Fundulus deep cells during early development. J. Cell. Sci. 13:721–726.PubMedGoogle Scholar
  41. Wayner, E.A., A. Garcia-Pardo, M.J. Humphries, J.A. McDonald, and W.G. Carter. 1989. Identification and characterization of the lymphocyte adhesion receptor for an alternative cell attachment domain (CS-1) in plasma fibronectin. J. Cell Biol. 109:1321–1330.PubMedCrossRefGoogle Scholar
  42. Wetts, R. and S.E. Fraser. 1989. Slow intermixing of cells during Xenopus embryogenesis contributes to the consistency of the fate map. Development 105:9–15.PubMedGoogle Scholar
  43. Winklbauer, R. 1988. Differential interaction of Xenopus embryonic cells with fibronectin in vitro. Dev. Biol. 130:175–183.PubMedCrossRefGoogle Scholar
  44. Winklbauer, R. 1990. Mesodermal cell migration during Xenopus gastrulation. Dev. Biol. 142:155–168.PubMedCrossRefGoogle Scholar
  45. Wollweber, L., R. Stracke, and U. Gothe. 1981. The use of a simple method to avoid cell shrinkage during SEM preparation. J. Microscopy 121:185–189.CrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1991

Authors and Affiliations

  • Douglas W. DeSimone
    • 1
  • Jim C. Smith
    • 2
  • James E. Howard
    • 2
  • David G. Ransom
    • 1
  • Karen Symes
    • 2
  1. 1.Department of Anatomy and Cell Biology and the Molecular Biology InstituteUniversity of Virginia Health Sciences CenterCharlottesvilleUSA
  2. 2.Laboratory of EmbryogenesisNational Institute for Medical ResearchLondonUK

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