Expression of cell adhesion molecule E-cadherin in Xenopus embryos begins at gastrulation and predominates in the ectoderm

The expression of the Ca2+-dependent epithelial cell adhesion molecule E-cadherin (also known as uvomorulin and L-CAM) in the early stages of embryonic development of Xenopus laevis was examined. E-Cadherin was identified in the Xenopus A6 epithelial cell line by antibody cross- reactivity and several biochemical characteristics. Four independent mAbs were generated against purified Xenopus E-cadherin. All four mAbs recognized the same polypeptides in A6 cells, adult epithelial tissues, and embryos. These mAbs inhibited the formation of cell contacts between A6 cells and stained the basolateral plasma membranes of A6 cells, hepatocytes, and alveolar epithelial cells. The time of E- cadherin expression in early Xenopus embryos was determined by immunoblotting. Unlike its expression in early mouse embryos, E- cadherin was not present in the eggs or early blastula of Xenopus laevis. These findings indicate that a different Ca2+-dependent cell adhesion molecule, perhaps another member of the cadherin gene family, is responsible for the Ca2+-dependent adhesion between cleavage stage Xenopus blastomeres. Detectable accumulation of E-cadherin started just before gastrulation at stage 9 1/2 and increased rapidly up to the end of gastrulation at stage 15. In stage 15 embryos, specific immunofluorescence staining of E-cadherin was discernible only in ectoderm, but not in mesoderm and endoderm. The ectoderm at this stage consists of two cell layers. The outer cell layer of ectoderm was stained intensely, and staining was localized to the basolateral plasma membrane of these cells. Lower levels of staining were observed in the inner cell layer of ectoderm. The coincidence of E-cadherin expression with the process of gastrulation and its restriction to the ectoderm indicate that it may play a role in the morphogenetic movements of gastrulation and resulting segregation of embryonic germ layers.

[1]  M. Takeichi,et al.  The cadherins: cell-cell adhesion molecules controlling animal morphogenesis. , 1988, Development.

[2]  H. Ploegh,et al.  Uvomorulin: a nonintegral membrane protein of early mouse embryo. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[3]  Philip L. Townes,et al.  Directed movements and selective adhesion of embryonic amphibian cells , 1955 .

[4]  S. D. de Laat,et al.  NEW MEMBRANE FORMATION DURING CYTOKINESIS IN NORMAL AND CYTOCHALASIN B-TREATED EGGS OF XENOPUS LAEVIS , 1973, The Journal of cell biology.

[5]  M. Takeichi,et al.  The calcium-dependent cell-cell adhesion system regulates inner cell mass formation and cell surface polarization in early mouse development , 1983, Cell.

[6]  G. Edelman,et al.  Ontogenetic expression of cell adhesion molecules: L-CAM is found in epithelia derived from the three primary germ layers. , 1984, Developmental biology.

[7]  B. Gumbiner,et al.  The role of the cell adhesion molecule uvomorulin in the formation and maintenance of the epithelial junctional complex , 1988, The Journal of cell biology.

[8]  J. Shih,et al.  The function and mechanism of convergent extension during gastrulation of Xenopus laevis. , 1985, Journal of embryology and experimental morphology.

[9]  G. Edelman,et al.  Expression sequences and distribution of two primary cell adhesion molecules during embryonic development of Xenopus laevis , 1987, The Journal of cell biology.

[10]  A. Nose,et al.  A novel cadherin cell adhesion molecule: its expression patterns associated with implantation and organogenesis of mouse embryos , 1986, The Journal of cell biology.

[11]  E. Jones,et al.  The development of animal cap cells in Xenopus: the effects of environment on the differentiation and the migration of grafted ectodermal cells , 1987 .

[12]  W. Schaffner,et al.  A rapid, sensitive, and specific method for the determination of protein in dilute solution. , 1973, Analytical biochemistry.

[13]  F. Jacob,et al.  Cell-cell interactions in early embryogenesis: A molecular approach to the role of calcium , 1981, Cell.

[14]  D. Vestweber,et al.  Some structural and functional aspects of the cell adhesion molecule uvomorulin. , 1984, Cell differentiation.

[15]  P. Hausen,et al.  Changes in the nuclear lamina composition during early development of Xenopus laevis , 1985, Cell.

[16]  Akinao Nose,et al.  Expressed recombinant cadherins mediate cell sorting in model systems , 1988, Cell.

[17]  R. Steinhardt,et al.  Global properties of the Xenopus blastula are mediated by a high-resistance epithelial seal☆ , 1986 .

[18]  M. Kirschner,et al.  Temporal and spatial regulation of fibronectin in early Xenopus development , 1984, Cell.

[19]  N. Peyriéras,et al.  Characterization of antigens recognized by monoclonal and polyclonal antibodies directed against uvomorulin. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[20]  G. Edelman,et al.  Sequence analysis of a cDNA clone encoding the liver cell adhesion molecule, L-CAM. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[21]  D. Vestweber,et al.  Expression and distribution of cell adhesion molecule uvomorulin in mouse preimplantation embryos. , 1987, Developmental biology.

[22]  R. Keller,et al.  Rearrangement of enveloping layer cells without disruption of the epithelial permeability barrier as a factor in Fundulus epiboly. , 1987, Developmental biology.

[23]  A. Nose,et al.  Cloning and expression of cDNA encoding a neural calcium-dependent cell adhesion molecule: its identity in the cadherin gene family , 1988, The Journal of cell biology.

[24]  D. Solter,et al.  Identification and purification of a cell surface glycoprotein mediating intercellular adhesion in embryonic and adult tissue , 1983, Cell.

[25]  M. Takeichi,et al.  A monoclonal antibody disrupting calcium-dependent cell-cell adhesion of brain tissues: possible role of its target antigen in animal pattern formation. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[26]  M. Takeichi,et al.  Calcium-dependent cell-cell adhesion molecules common to hepatocytes and teratocarcinoma stem cells , 1983, The Journal of cell biology.

[27]  M. Ringwald,et al.  The structure of cell adhesion molecule uvomorulin. Insights into the molecular mechanism of Ca2+‐dependent cell adhesion. , 1987, The EMBO journal.

[28]  M. Kirschner,et al.  A major developmental transition in early xenopus embryos: I. characterization and timing of cellular changes at the midblastula stage , 1982, Cell.

[29]  C. Damsky,et al.  Developmentally regulated expression of the cell-cell adhesion glycoprotein cell-CAM 120/80 in peri-implantation mouse embryos and extraembryonic membranes. , 1986, Developmental biology.

[30]  K. Shiokawa,et al.  Cell to Cell Adhesion Systems in Xenopus laevis, the South African Clawed Frog I. Detection of Ca2+ Dependent and Independent Adhesion Systems in Adult and Embryonic Cells , 1986 .

[31]  J. Gerhart,et al.  The timing of early developmental events in Xenopus , 1985 .

[32]  G. Warren,et al.  A monoclonal antibody against a 135‐K Golgi membrane protein. , 1982, The EMBO journal.

[33]  E. Adamson,et al.  The synthesis and storage of histones during the oogenesis of Xenopus laevis. , 1977, Developmental biology.

[34]  F. Jacob,et al.  A cell surface glycoprotein involved in the compaction of embryonal carcinoma cells and cleavage stage embryos , 1980, Cell.

[35]  P. Singal,et al.  An ultrastructural study of the first cleavage of Xenopus embryos. , 1974, Journal of ultrastructure research.

[36]  I. Dawid,et al.  Xenopus laevis in developmental and molecular biology. , 1988, Science.

[37]  G. Edelman,et al.  Characterization of L-CAM, a major cell adhesion molecule from embryonic liver cells. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[38]  W. Birchmeier,et al.  Dissociation of Madin-Darby canine kidney epithelial cells by the monoclonal antibody anti-arc-1: mechanistic aspects and identification of the antigen as a component related to uvomorulin , 1985, The Journal of cell biology.

[39]  Stephen P. A. Brown,et al.  Monoclonal antibody technology. , 1986, American journal of hospital pharmacy.

[40]  C. Bordier Phase separation of integral membrane proteins in Triton X-114 solution. , 1981, The Journal of biological chemistry.

[41]  B. Gumbiner,et al.  A functional assay for proteins involved in establishing an epithelial occluding barrier: identification of a uvomorulin-like polypeptide , 1986, The Journal of cell biology.

[42]  N. Gilula,et al.  Antibodies to gap-junctional protein selectively disrupt junctional communication in the early amphibian embryo , 1984, Nature.

[43]  T. Tajima,et al.  Cell to cell adhesion systems in Xenopus laevis, the South African clawed toad. II: Monoclonal antibody against a novel Ca2+-dependent cell-cell adhesion glycoprotein on amphibian cells. , 1988, Cell differentiation.

[44]  J. Murphy,et al.  ROLE OF Na+, K+‐ATPase IN EARLY EMBRYONIC DEVELOPMENT * , 1974, Annals of the New York Academy of Sciences.

[45]  D. S. Phelps Electrophoretic transfer of proteins from fixed and stained gels. , 1984, Analytical biochemistry.

[46]  J. Gerhart,et al.  Region-specific cell activities in amphibian gastrulation. , 1986, Annual review of cell biology.

[47]  D. Vestweber,et al.  Cell-adhesion molecule uvomorulin is localized in the intermediate junctions of adult intestinal epithelial cells , 1985, The Journal of cell biology.

[48]  M. Takeichi,et al.  Spatial and temporal expression pattern of N-cadherin cell adhesion molecules correlated with morphogenetic processes of chicken embryos. , 1987, Developmental biology.

[49]  M. Takeichi,et al.  Expression of N-cadherin adhesion molecules associated with early morphogenetic events in chick development , 1986, Nature.