title: Phosphorylation of ezrin on threonine T567 plays a crucial role during compaction in the mouse early embryo.
abstract: The preimplantation development of the mouse embryo leads to the divergence of the first two cell lineages, the inner cell mass and the trophectoderm. The formation of a microvillus pole during compaction at the eight-cell stage and its asymmetric inheritance during mitosis are key events in the emergence of these two cell populations. Ezrin, a member of the ERM protein family, seems to be involved in the formation and stabilization of this apical microvillus pole. To further characterize its function in early development, we mutated the key residue T567, which was reported to be essential for regulation of ezrin function through phosphorylation. Here, we show that expression of ezrin mutants in which the COOH-terminal threonine T567 was replaced by an aspartate (to mimic a phosphorylated residue; T567D) or by an alanine (to avoid phosphorylation; T567A) interferes with E-cadherin function and disrupts the first morphogenetic events of development: compaction and cavitation. The active mutant ezrin-T567D induces the formation of numerous and abnormally long microvilli at the surface of blastomeres. Moreover, it localizes all around the cell cortex and inhibits cell-cell adhesion and cell polarization at the eight-cell stage. During the following stages, only half of the embryos are able to compact and finally to cavitate. In those embryos, the amount of ezrin-T567D decreases in the basolateral areas, while the proportion of adherens junctions increases. The reverse inactive mutant ezrin-T567A is mainly cytoplasmic and does not perturb compaction at the eight-cell stage. However, at the 16-cell stage, it relocalizes at the basolateral cortex, leading to a strong decrease in the surface of adherens junctions, and finally, embryos abort development. Our results show that ezrin is directly involved in the formation of microvilli in the early mouse embryo. Moreover, they indicate that maintenance of ezrin in basolateral areas prevents microvilli breakdown and inhibits the formation of normal cell-cell contacts mediated by E-cadherin, thereby impairing blastomeres polarization and morphogenesis of the blastocyst.
authors.affiliation: Laboratoire de Biologie Cellulaire du Développement, UMR7622, CNRS-Université Pierre et Marie Curie, 9 Quai St-Bernard, 75252 Paris cedex 05, France. nicolas.dard@bc.biol.ethz.ch
authors.forename: Nicolas
authors.initials: N
authors.lastname: Dard
authors.forename: Sophie
authors.initials: S
authors.lastname: Louvet-Vallée
authors.forename: Angélica
authors.initials: A
authors.lastname: Santa-Maria
authors.forename: Bernard
authors.initials: B
authors.lastname: Maro
mesh_terms: D019943:Amino Acid Substitution / Q000235:genetics / Q000502:physiology*; D000818:Animals; D002448:Cell Adhesion / Q000235:genetics / Q000502:physiology; D016764:Cell Polarity / Q000235:genetics / Q000502:physiology; D003598:Cytoskeletal Proteins; D017931:DNA Primers; D005260:Female; D018507:Gene Expression Regulation, Developmental; D049452:Green Fluorescent Proteins; D007150:Immunohistochemistry; D008164:Luminescent Proteins; D051379:Mice / Q000196:embryology*; D008845:Microinjections; D008854:Microscopy, Electron; D018715:Microscopy, Video; D008871:Microvilli / Q000235:genetics / Q000502:physiology*; D009028:Morula / Q000502:physiology / Q000648:ultrastructure*; D016297:Mutagenesis, Site-Directed; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D010766:Phosphorylation; D012333:RNA, Messenger / Q000378:metabolism*; D000099149:Ezrin
chemical_list: D003598:Cytoskeletal Proteins; D017931:DNA Primers; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D010750:Phosphoproteins; D012333:RNA, Messenger; D000099149:Ezrin