Transiently reorganized microtubules are essential for zippering during dorsal closure in Drosophila melanogaster.

There is emerging evidence that microtubules in nondividing cells can be employed to remodel the intracellular space. Here, we demonstrate an essential role for microtubules in dorsal closure, which occurs toward the end of Drosophila melanogaster embryogenesis. Dorsal closure is a morphogenetic process similar to wound healing, whereby a gap in the epithelium is closed through the coordinated action of different cell types. Surprisingly, this complex process requires microtubule function exclusively in epithelial cells and only for the last step, the zippering, which seals the gap. Preceding zippering, the epithelial microtubules reorganize to attain an unusual spatial distribution, which we describe with subcellular resolution in the intact, living organism. We provide a clearly defined example where cells of a developing organism transiently reorganize their microtubules to fulfill a specialized morphogenetic task.

[1]  P. Rørth,et al.  Invasive cell migration is initiated by guided growth of long cellular extensions , 2002, Nature Cell Biology.

[2]  K. Edwards,et al.  GFP-moesin illuminates actin cytoskeleton dynamics in living tissue and demonstrates cell shape changes during morphogenesis in Drosophila. , 1997, Developmental biology.

[3]  M. Welte,et al.  Bidirectional Transport along Microtubules , 2004, Current Biology.

[4]  A. Hall,et al.  Integrin-Mediated Activation of Cdc42 Controls Cell Polarity in Migrating Astrocytes through PKCζ , 2001, Cell.

[5]  D. Strutt Asymmetric localization of frizzled and the establishment of cell polarity in the Drosophila wing. , 2001, Molecular cell.

[6]  G. Gundersen,et al.  Low concentrations of nocodazole interfere with fibroblast locomotion without significantly affecting microtubule level: implications for the role of dynamic microtubules in cell locomotion. , 1995, Journal of cell science.

[7]  S. Bagrodia,et al.  The Dbl-related Protein, Lfc, Localizes to Microtubules and Mediates the Activation of Rac Signaling Pathways in Cells* , 1999, The Journal of Biological Chemistry.

[8]  K. Broadie,et al.  The Hereditary Spastic Paraplegia Gene, spastin, Regulates Microtubule Stability to Modulate Synaptic Structure and Function , 2004, Current Biology.

[9]  R. Pepperkok,et al.  Spectral imaging and its applications in live cell microscopy , 2003, FEBS letters.

[10]  P. Nurse,et al.  tea1 and the Microtubular Cytoskeleton Are Important for Generating Global Spatial Order within the Fission Yeast Cell , 1997, Cell.

[11]  Paul Martin,et al.  The small GTPase Rac plays multiple roles in epithelial sheet fusion--dynamic studies of Drosophila dorsal closure. , 2005, Developmental biology.

[12]  A. Bershadsky,et al.  Microtubule-dependent control of cell shape and pseudopodial activity is inhibited by the antibody to kinesin motor domain , 1993, The Journal of cell biology.

[13]  M. Martinez-Campos,et al.  The Drosophila pericentrin-like protein is essential for cilia/flagella function, but appears to be dispensable for mitosis , 2004, The Journal of cell biology.

[14]  Y. Zheng,et al.  Cloning and Characterization of GEF-H1, a Microtubule-associated Guanine Nucleotide Exchange Factor for Rac and Rho GTPases* , 1998, The Journal of Biological Chemistry.

[15]  E. Salmon,et al.  Microtubule growth activates Rac1 to promote lamellipodial protrusion in fibroblasts , 1999, Nature Cell Biology.

[16]  L. Lim,et al.  A Drosophila homolog of the Rac- and Cdc42-activated serine/threonine kinase PAK is a potential focal adhesion and focal complex protein that colocalizes with dynamic actin structures , 1996, Molecular and cellular biology.

[17]  V. Doye,et al.  A Mechanism for Nuclear Positioning in Fission Yeast Based on Microtubule Pushing , 2001, The Journal of cell biology.

[18]  G. C. Rogers,et al.  Drosophila EB1 is important for proper assembly, dynamics, and positioning of the mitotic spindle , 2002, The Journal of cell biology.

[19]  K. Hahn,et al.  Activation of Endogenous Cdc42 Visualized in Living Cells , 2004, Science.

[20]  S. Étienne-Manneville Actin and Microtubules in Cell Motility: Which One is in Control? , 2004, Traffic.

[21]  Marileen Dogterom,et al.  Force generation by dynamic microtubules. , 2005, Current opinion in cell biology.

[22]  A. Martinez-Arias,et al.  Dynamic actin-based epithelial adhesion and cell matching during Drosophila dorsal closure , 2000, Current Biology.

[23]  J. A. Kaltschmidt,et al.  Planar polarity and actin dynamics in the epidermis of Drosophila , 2002, Nature Cell Biology.

[24]  I. Vernos,et al.  The Mitotic Spindle: A Self-Made Machine , 2001, Science.

[25]  G. C. Rogers,et al.  Functionally distinct kinesin-13 family members cooperate to regulate microtubule dynamics during interphase , 2005, Nature Cell Biology.

[26]  M. Peifer,et al.  Drosophila APC2 and Armadillo participate in tethering mitotic spindles to cortical actin , 2001, Nature Cell Biology.

[27]  C. Hoogenraad,et al.  Microtubule plus-end-tracking proteins: mechanisms and functions. , 2005, Current opinion in cell biology.

[28]  T. Mitchison,et al.  Microtubule polymerization dynamics. , 1997, Annual review of cell and developmental biology.

[29]  C. Turck,et al.  Drosophila RhoGEF2 Associates with Microtubule Plus Ends in an EB1-Dependent Manner , 2004, Current Biology.

[30]  James Q. Zheng,et al.  Cytoskeletal dynamics underlying collateral membrane protrusions induced by neurotrophins in cultured Xenopus embryonic neurons. , 2003, Journal of neurobiology.

[31]  Ilan Davis,et al.  Drosophila wingless and Pair-Rule Transcripts Localize Apically by Dynein-Mediated Transport of RNA Particles , 2001, Cell.

[32]  P. Nurse,et al.  CLIP170-like tip1p Spatially Organizes Microtubular Dynamics in Fission Yeast , 2000, Cell.

[33]  Wayne L. Rickoll,et al.  Multiple Forces Contribute to Cell Sheet Morphogenesis for Dorsal Closure in Drosophila , 2000, The Journal of cell biology.

[34]  Paul Martin,et al.  Dynamic analysis of dorsal closure in Drosophila: from genetics to cell biology. , 2002, Developmental cell.

[35]  P. Nurse,et al.  The Kinesin Klp2 Mediates Polarization of Interphase Microtubules in Fission Yeast , 2005, Science.

[36]  Alfonso Martinez Arias,et al.  Armadillo/β-catenin-dependent Wnt signalling is required for the polarisation of epidermal cells during dorsal closure in Drosophila , 2004, Development.

[37]  A. Spradling,et al.  The fusome organizes the microtubule network during oocyte differentiation in Drosophila. , 2000, Development.

[38]  D. Berdnik,et al.  Drosophila Aurora-A Is Required for Centrosome Maturation and Actin-Dependent Asymmetric Protein Localization during Mitosis , 2002, Current Biology.

[39]  F. Turner,et al.  The centrosome is a dynamic structure that ejects PCM flares , 2002, Journal of Cell Science.

[40]  Paul Martin,et al.  Mechanisms of epithelial fusion and repair , 2001, Nature Cell Biology.

[41]  B. Alberts,et al.  Reorganization of the cytoskeleton during Drosophila oogenesis: implications for axis specification and intercellular transport. , 1992, Development.

[42]  P. Rørth Gal4 in the Drosophila female germline , 1998, Mechanisms of Development.

[43]  H. Jasper,et al.  JNK signaling coordinates integrin and actin functions during Drosophila embryogenesis , 2006, Developmental dynamics : an official publication of the American Association of Anatomists.

[44]  Anthony A. Hyman,et al.  Polarity controls forces governing asymmetric spindle positioning in the Caenorhabditis elegans embryo , 2001, Nature.

[45]  Y. Koh,et al.  Regulation of DLG Localization at Synapses by CaMKII-Dependent Phosphorylation , 1999, Cell.

[46]  K. Oegema,et al.  The minus end in sight , 2003, Current Biology.

[47]  E. Karsenti,et al.  Microtubule‐dependent transport and organization of sarcomeric myosin during skeletal muscle differentiation , 2005, The EMBO journal.

[48]  Two‐color GFP imaging demonstrates cell‐autonomy of GAL4‐driven RNA interference in drosophila , 2002, Genesis.

[49]  Steven P Gross,et al.  Developmental Regulation of Vesicle Transport in Drosophila Embryos: Forces and Kinetics , 1998, Cell.

[50]  James Q. Zheng,et al.  Growth Cone Turning Induced by Direct Local Modification of Microtubule Dynamics , 2002, The Journal of Neuroscience.

[51]  A. Ephrussi,et al.  Axis formation during Drosophila oogenesis. , 2001, Current opinion in genetics & development.

[52]  K. Zinn,et al.  Drosophila Spastin Regulates Synaptic Microtubule Networks and Is Required for Normal Motor Function , 2004, PLoS biology.

[53]  G. Edwards,et al.  Forces for Morphogenesis Investigated with Laser Microsurgery and Quantitative Modeling , 2003, Science.