Endocytosis Is Required for Efficient Apical Constriction during Xenopus Gastrulation

Coordinated apical constriction (AC) in epithelial sheets drives tissue invagination [1, 2] and is required for diverse morphogenetic movements such as gastrulation [3], neurulation [4, 5], and organogenesis [6]. We showed previously that actomyosin contractility drives AC in Xenopus laevis bottle cells [7]; however, it remained unclear whether it does so in concert with other processes. Here we report that endocytosis-driven membrane remodeling is required for efficient AC. We found endosomes exclusively in bottle cells in the early gastrula. Disrupting endocytosis with dominant-negative dynamin or rab5 perturbed AC, with a significant decrease in constriction rate late in the process, suggesting that endocytosis operates downstream of actomyosin contractility to remove excess membrane. Additionally, disrupting endocytosis during neurulation inhibits AC in hingepoint cells, resulting in neural tube closure defects. Thus, membrane remodeling during AC could be a general mechanism to achieve efficient invagination in embryos.

[1]  E. Schierenberg,et al.  The cellular anatomy of embryos of the nematode Caenorhabditis elegans. Analysis and reconstruction of serial section electron micrographs. , 1978, Developmental biology.

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

[3]  T. Kreis,et al.  Translocation and clustering of endosomes and lysosomes depends on microtubules , 1987, The Journal of cell biology.

[4]  J. Hardin,et al.  Bottle cells are required for the initiation of primary invagination in the sea urchin embryo. , 1998, Developmental biology.

[5]  S. Schmid,et al.  Induction of mutant dynamin specifically blocks endocytic coated vesicle formation , 1994, The Journal of cell biology.

[6]  V. Tsarouhas,et al.  Sequential pulses of apical epithelial secretion and endocytosis drive airway maturation in Drosophila. , 2007, Developmental cell.

[7]  L. Wolpert,et al.  The amniote primitive streak is defined by epithelial cell intercalation before gastrulation , 2007, Nature.

[8]  K. Miyasaka,et al.  Daam1 regulates the endocytosis of EphB during the convergent extension of the zebrafish notochord , 2007, Proceedings of the National Academy of Sciences.

[9]  W. H. Lewis,et al.  Mechanics of invagination , 1947, The Anatomical record.

[10]  J. Wallingford,et al.  Neural tube closure requires Dishevelled-dependent convergent extension of the midline , 2002, Development.

[11]  R. Keller,et al.  Surface mesoderm in Xenopus: a revision of the stage 10 fate map , 1997, Development Genes and Evolution.

[12]  D. Andrew,et al.  Ribbon modulates apical membrane during tube elongation through Crumbs and Moesin. , 2008, Developmental biology.

[13]  Gun-Hwa Kim,et al.  Ryk cooperates with Frizzled 7 to promote Wnt11-mediated endocytosis and is essential for Xenopus laevis convergent extension movements , 2008, The Journal of cell biology.

[14]  A. Copp.  Neurulation in the cranial region – normal and abnormal , 2005, Journal of anatomy.

[15]  James C. Smith,et al.  Rab5-mediated endocytosis of activin is not required for gene activation or long-range signalling in Xenopus , 2009, Development.

[16]  D. Andrew,et al.  Organ shape in the Drosophila salivary gland is controlled by regulated, sequential internalization of the primordia. , 2000, Development.

[17]  Bob Goldstein,et al.  Mechanisms of cell positioning during C. elegans gastrulation , 2003, Development.

[18]  G. Schoenwolf,et al.  Quantitative analyses of changes in cell shapes during bending of the avian neural plate. , 1984, Developmental biology.

[19]  Nancy Papalopulu,et al.  Defining a large set of full-length clones from a Xenopus tropicalis EST project. , 2004, Developmental biology.

[20]  Anna V. Taubenberger,et al.  Wnt11 functions in gastrulation by controlling cell cohesion through Rab5c and E-cadherin. , 2005, Developmental cell.

[21]  R. Keller,et al.  Epithelial type, ingression, blastopore architecture and the evolution of chordate mesoderm morphogenesis. , 2008, Journal of experimental zoology. Part B, Molecular and developmental evolution.

[22]  T. Lecuit,et al.  Trafficking through Rab11 Endosomes Is Required for Cellularization during Drosophila Embryogenesis , 2003, Current Biology.

[23]  R. Keller,et al.  An experimental analysis of the role of bottle cells and the deep marginal zone in gastrulation of Xenopus laevis. , 1981, The Journal of experimental zoology.

[24]  P. L. Campbell,et al.  EEA1, an Early Endosome-Associated Protein. , 1995, The Journal of Biological Chemistry.

[25]  R. Harland,et al.  Actomyosin contractility and microtubules drive apical constriction in Xenopus bottle cells. , 2007, Developmental biology.

[26]  P. Stahl,et al.  Structure-function relationship of the small GTPase rab5. , 1993, The Journal of biological chemistry.

[27]  J. Gurdon,et al.  Morphogen gradient interpretation by a regulated trafficking step during ligand-receptor transduction. , 2005, Genes & development.

[28]  J. Wallingford,et al.  Shroom Induces Apical Constriction and Is Required for Hingepoint Formation during Neural Tube Closure , 2003, Current Biology.

[29]  H. Fares,et al.  Genome-wide analysis identifies a general requirement for polarity proteins in endocytic traffic , 2007, Nature Cell Biology.

[30]  M. Zerial,et al.  Localization of low molecular weight GTP binding proteins to exocytic and endocytic compartments , 1990, Cell.

[31]  B. Burnside,et al.  Microtubules and microfilaments in newt neuralation. , 1971, Developmental biology.

[32]  Ray Keller,et al.  How we are shaped: the biomechanics of gastrulation. , 2003, Differentiation; research in biological diversity.

[33]  M. Llimargas,et al.  Modulation of intracellular trafficking regulates cell intercalation in the Drosophila trachea , 2008, Nature Cell Biology.

[34]  D. Bilder,et al.  Endocytic control of epithelial polarity and proliferation in Drosophila , 2005, Nature Cell Biology.

[35]  P. Alberch,et al.  The mechanical basis of morphogenesis. I. Epithelial folding and invagination. , 1981, Developmental biology.

[36]  R. Keller,et al.  Pattern and morphogenesis of presumptive superficial mesoderm in two closely related species, Xenopus laevis and Xenopus tropicalis. , 2004, Developmental biology.

[37]  T. Kurth,et al.  Bottle cell formation in relation to mesodermal patterning in the Xenopus embryo , 2000, Mechanisms of Development.

[38]  Ken W. Y. Cho,et al.  TGF-beta signaling-mediated morphogenesis: modulation of cell adhesion via cadherin endocytosis. , 2007, Genes & development.

[39]  J. Holtfreter.  A study of the mechanics of gastrulation. Part I , 1943 .