Integrin α5β1 and Fibronectin Regulate Polarized Cell Protrusions Required for Xenopus Convergence and Extension

Summary Background Integrin recognition of fibronectin is required for normal gastrulation including the mediolateral cell intercalation behaviors that drive convergent extension and the elongation of the frog dorsal axis; however, the cellular and molecular mechanisms involved are unclear. Results We report that depletion of fibronectin with antisense morpholinos blocks both convergent extension and mediolateral protrusive behaviors in explant preparations. Both chronic depletion of fibronectin and acute disruptions of integrin α 5 β 1 binding to fibronectin increases the frequency and randomizes the orientation of polarized cellular protrusions, suggesting that integrin-fibronectin interactions normally repress frequent random protrusions in favor of fewer mediolaterally oriented ones. In the absence of integrin α 5 β 1 binding to fibronectin, convergence movements still occur but result in convergent thickening instead of convergent extension. Conclusions These findings support a role for integrin signaling in regulating the protrusive activity that drives axial extension. We hypothesize that the planar spatial arrangement of the fibrillar fibronectin matrix, which delineates tissue compartments within the embryo, is critical for promoting productive oriented protrusions in intercalating cells.

[1]  R. Hynes,et al.  Overlapping and independent functions of fibronectin receptor integrins in early mesodermal development. , 1999, Developmental biology.

[2]  R. Hynes,et al.  The dynamic dialogue between cells and matrices: implications of fibronectin's elasticity. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[3]  R. Hynes,et al.  Identification and characterization of alternatively spliced fibronectin mRNAs expressed in early Xenopus embryos. , 1992, Developmental biology.

[4]  J. Taipale,et al.  Latent transforming growth factor-beta binding proteins (LTBPs)--structural extracellular matrix proteins for targeting TGF-beta action. , 1999, Cytokine & growth factor reviews.

[5]  R. Winklbauer,et al.  Vegetal rotation, a new gastrulation movement involved in the internalization of the mesoderm and endoderm in Xenopus. , 1999, Development.

[6]  Ray Keller,et al.  Mesendoderm Extension and Mantle Closure in Xenopus laevis Gastrulation: Combined Roles for Integrin α5β1, Fibronectin, and Tissue Geometry , 2002 .

[7]  J. Schwarzbauer,et al.  Modulatory roles for integrin activation and the synergy site of fibronectin during matrix assembly. , 1997, Molecular biology of the cell.

[8]  B. Kay,et al.  Xenopus laevis : practical uses in cell and molecular biology , 1991 .

[9]  M. Marsden,et al.  Regulation of cell polarity, radial intercalation and epiboly in Xenopus: novel roles for integrin and fibronectin. , 2001, Development.

[10]  Scott E. Fraser,et al.  Dishevelled controls cell polarity during Xenopus gastrulation , 2000, Nature.

[11]  R. Keller,et al.  Assembly and remodeling of the fibrillar fibronectin extracellular matrix during gastrulation and neurulation in Xenopus laevis , 2004, Developmental dynamics : an official publication of the American Association of Anatomists.

[12]  D. O'Leary,et al.  Labeling Neural Cells Using Adenoviral Gene Transfer of Membrane-Targeted GFP , 1996, Neuron.

[13]  D. DeSimone,et al.  Xenopus embryonic cell adhesion to fibronectin: position-specific activation of RGD/synergy site-dependent migratory behavior at gastrulation , 1996, The Journal of cell biology.

[14]  J. Shih,et al.  Cell motility driving mediolateral intercalation in explants of Xenopus laevis. , 1992, Development.

[15]  Lance A Davidson,et al.  Patterning and tissue movements in a novel explant preparation of the marginal zone of Xenopus laevis. , 2004, Gene expression patterns : GEP.

[16]  Douglas W. DeSimone,et al.  Integrin-ECM Interactions Regulate Cadherin-Dependent Cell Adhesion and Are Required for Convergent Extension in Xenopus , 2003, Current Biology.

[17]  Martin A. Schwartz,et al.  Networks and crosstalk: integrin signalling spreads , 2002, Nature Cell Biology.

[18]  C. Harley,et al.  In situ analysis of changes in telomere size during replicative aging and cell transformation , 1996, The Journal of cell biology.

[19]  P. Skoglund,et al.  Mechanisms of convergence and extension by cell intercalation. , 2000, Philosophical transactions of the Royal Society of London. Series B, Biological sciences.

[20]  J. Schwarzbauer,et al.  Fibronectin matrix composition and organization can regulate cell migration during amphibian development , 2000, Mechanisms of Development.

[21]  M. Hatzfeld The p120 family of cell adhesion molecules. , 2005, European journal of cell biology.

[22]  Scott E Fraser,et al.  Convergent extension: the molecular control of polarized cell movement during embryonic development. , 2002, Developmental Cell.

[23]  T. Darribère,et al.  Fibronectin-rich fibrillar extracellular matrix controls cell migration during amphibian gastrulation. , 1990, The International journal of developmental biology.

[24]  R. Hynes,et al.  Embryonic mesodermal defects in 5 integrin-deficient mice , 1996 .

[25]  M. Koehl,et al.  The dorsal involuting marginal zone stiffens anisotropically during its convergent extension in the gastrula of Xenopus laevis. , 1995, Development.

[26]  L. Solnica-Krezel Conserved Patterns of Cell Movements during Vertebrate Gastrulation , 2005, Current Biology.

[27]  J. Thiery,et al.  Evidence for the role of fibronectin in amphibian gastrulation. , 1985, Journal of embryology and experimental morphology.

[28]  D. DeSimone,et al.  Molecular analysis and developmental expression of the focal adhesion kinase pp125FAK in Xenopus laevis. , 1995, Developmental biology.

[29]  Ray Keller,et al.  Planar Cell Polarity Genes Regulate Polarized Extracellular Matrix Deposition during Frog Gastrulation , 2005, Current Biology.

[30]  C. Heisenberg,et al.  Gastrulation dynamics: cells move into focus. , 2004, Trends in cell biology.

[31]  C. Stern Gastrulation : from cells to embryo , 2004 .

[32]  Richard Milner,et al.  The integrin family of cell adhesion molecules has multiple functions within the CNS , 2002, Journal of neuroscience research.

[33]  J. Shih,et al.  Patterns of cell motility in the organizer and dorsal mesoderm of Xenopus laevis. , 1992, Development.

[34]  R. Harland,et al.  Early development of Xenopus laevis : a laboratory manual , 2000 .

[35]  P. Skoglund,et al.  The midline (notochord and notoplate) patterns the cell motility underlying convergence and extension of the Xenopus neural plate. , 2003, Developmental biology.

[36]  D. DeSimone,et al.  Integrin-dependent adhesive activity is spatially controlled by inductive signals at gastrulation. , 1996, Development.

[37]  Ray Keller,et al.  Shaping the Vertebrate Body Plan by Polarized Embryonic Cell Movements , 2002, Science.

[38]  R. Hynes,et al.  Antibodies to the conserved cytoplasmic domain of the integrin beta 1 subunit react with proteins in vertebrates, invertebrates, and fungi , 1988, The Journal of cell biology.

[39]  T. Joos,et al.  Development and control of tissue separation at gastrulation in Xenopus. , 2000, Developmental biology.

[40]  R. Hynes,et al.  Embryonic mesodermal defects in alpha 5 integrin-deficient mice. , 1993, Development.

[41]  R. Keller,et al.  The Cellular Basis of Gastrulation in Xenopus laevis: Active, Postinvolution Convergence and Extension by Mediolateral Interdigitation , 1984 .

[42]  J. Thiery,et al.  Prevention of gastrulation but not neurulation by antibodies to fibronectin in amphibian embryos , 1984, Nature.

[43]  R. Hynes,et al.  Defects in mesoderm, neural tube and vascular development in mouse embryos lacking fibronectin. , 1993, Development.

[44]  S. Pizer,et al.  The Image Processing Handbook , 1994 .

[45]  R. Keller,et al.  Induction of neuronal differentiation by planar signals in Xenopus embryos , 1993, Developmental dynamics : an official publication of the American Association of Anatomists.

[46]  R. Winklbauer,et al.  Antero-posterior tissue polarity links mesoderm convergent extension to axial patterning , 2004, Nature.

[47]  L. Davidson,et al.  Neural tube closure in Xenopus laevis involves medial migration, directed protrusive activity, cell intercalation and convergent extension. , 1999, Development.

[48]  M. Tessier-Lavigne,et al.  Recognition of the Neural Chemoattractant Netrin-1 by Integrins α6β4 and α3β1 Regulates Epithelial Cell Adhesion and Migration , 2003 .

[49]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[50]  E. Ruoslahti,et al.  Regulation of the fibronectin receptor affinity by divalent cations. , 1988, The Journal of biological chemistry.

[51]  G. Bazzoni,et al.  Divalent Cations and Ligands Induce Conformational Changes That Are Highly Divergent among β1 Integrins* , 1998, The Journal of Biological Chemistry.

[52]  M. Humphries,et al.  Regulation of integrin alpha 5 beta 1-fibronectin interactions by divalent cations. Evidence for distinct classes of binding sites for Mn2+, Mg2+, and Ca2+. , 1995, The Journal of biological chemistry.

[53]  B. Gumbiner,et al.  Analysis of C-cadherin Regulation during Tissue Morphogenesis with an Activating Antibody , 1999, The Journal of cell biology.

[54]  R. Keller,et al.  The cellular basis of epiboly: an SEM study of deep-cell rearrangement during gastrulation in Xenopus laevis. , 1980, Journal of embryology and experimental morphology.

[55]  Jeff Hardin,et al.  Models of morphogenesis: the mechanisms and mechanics of cell rearrangement. , 2004, Current opinion in genetics & development.