Actin nucleator Arp2/3 complex is essential for mouse preimplantation embryo development.

The Arp2/3 complex is a critical actin nucleator, which promotes actin assembly and is widely involved in a diverse range of actin-related processes such as cell locomotion, phagocytosis and the establishment of cell polarity. Previous studies showed that the Arp2/3 complex regulates spindle migration and asymmetric division during mouse oocyte maturation; however, the role of the Arp2/3 complex in early mouse embryo development is still unknown. The results of the present study show that the Arp2/3 complex is critical for cytokinesis during mouse embryo development. The Arp2/3 complex was concentrated at the cortex of each cell at the 2- to 8-cell stage and the peripheral areas of the morula and blastocyst. Inhibition of the Arp2/3 complex by the specific inhibitor CK666 at the zygote stage caused a failure in cell division; mouse embryos failed to undergo compaction and lost apical-basal polarity. The actin level decreased in the CK666-treated group, and two or more nuclei were observed within a single cell, indicating a failure of cell division. Addition of CK666 at the 8-cell stage caused a failure of blastocyst formation, and CDX2 staining confirmed the loss of embryo polarity and the failure of trophectoderm and inner cell mass formation. Taken together, these data suggest that the Arp2/3 complex may regulate mouse embryo development via its effect on cell division.

[1]  GM130: New insights into oocyte asymmetric division , 2011, Cell cycle.

[2]  T. Gotow,et al.  Sleeping Beauty Transposon-Based Phenotypic Analysis of Mice: Lack of Arpc3 Results in Defective Trophoblast Outgrowth , 2006, Molecular and Cellular Biology.

[3]  X. Cui,et al.  WAVE2 regulates meiotic spindle stability, peripheral positioning and polar body emission in mouse oocytes , 2011, Cell cycle.

[4]  Nam-Hyung Kim,et al.  JMY is required for asymmetric division and cytokinesis in mouse oocytes. , 2011, Molecular human reproduction.

[5]  V. Moreau,et al.  The yeast actin-related protein Arp2p is required for the internalization step of endocytosis. , 1997, Molecular biology of the cell.

[6]  Y. Marikawa,et al.  Establishment of trophectoderm and inner cell mass lineages in the mouse embryo , 2009, Molecular reproduction and development.

[7]  J. Rossant Lineage development and polar asymmetries in the peri-implantation mouse blastocyst. , 2004, Seminars in cell & developmental biology.

[8]  H. Riezman,et al.  Functional interactions between the p35 subunit of the Arp2/3 complex and calmodulin in yeast. , 2000, Molecular biology of the cell.

[9]  T. Pollard Mechanics of cytokinesis in eukaryotes. , 2010, Current opinion in cell biology.

[10]  Matthew D. Welch,et al.  A nucleator arms race: cellular control of actin assembly , 2010, Nature Reviews Molecular Cell Biology.

[11]  M. Kirschner,et al.  The Arp2/3 complex mediates actin polymerization induced by the small GTP-binding protein Cdc42. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[12]  X. Cui,et al.  Arp2/3 Complex Regulates Asymmetric Division and Cytokinesis in Mouse Oocytes , 2011, PloS one.

[13]  B. Goldstein,et al.  In vivo roles for Arp2/3 in cortical actin organization during C. elegans gastrulation , 2009, Journal of Cell Science.

[14]  Brian D. Slaughter,et al.  Dynamic maintenance of asymmetric meiotic spindle position through Arp2/3-complex-driven cytoplasmic streaming in mouse oocytes , 2011, Nature Cell Biology.

[15]  Fred Chang,et al.  Actin dynamics in the contractile ring during cytokinesis in fission yeast , 2002, Nature.

[16]  J. Segall,et al.  The F-actin side binding activity of the Arp2/3 complex is essential for actin nucleation and lamellipod extension , 2001, Current Biology.

[17]  B. Baum,et al.  Cdc42, Par6, and aPKC Regulate Arp2/3-Mediated Endocytosis to Control Local Adherens Junction Stability , 2008, Current Biology.

[18]  R. Vale,et al.  Molecular requirements for actin-based lamella formation in Drosophila S2 cells , 2003, The Journal of cell biology.

[19]  Wael Tadros,et al.  The maternal-to-zygotic transition: a play in two acts , 2009, Development.

[20]  J. Segall,et al.  Molecular mechanisms of invadopodium formation , 2005, The Journal of cell biology.

[21]  S. Coonrod,et al.  Use of proteomics to identify highly abundant maternal factors that drive the egg-to-embryo transition. , 2010, Reproduction.

[22]  I. Cameron,et al.  PKC signalling regulates tight junction membrane assembly in the pre-implantation mouse embryo. , 2004, Reproduction.

[23]  Janet Rossant,et al.  Disorganized epithelial polarity and excess trophectoderm cell fate in preimplantation embryos lacking E-cadherin , 2010, Development.

[24]  T. Pollard,et al.  Characterization of two classes of small molecule inhibitors of Arp2/3 complex , 2009, Nature.

[25]  Klemens Rottner,et al.  Filopodia formation in the absence of functional WAVE- and Arp2/3-complexes. , 2006, Molecular biology of the cell.

[26]  W. Mohler,et al.  The branched actin nucleator Arp2/3 promotes nuclear migrations and cell polarity in the C. elegans zygote. , 2011, Developmental biology.

[27]  M. Kirschner,et al.  The Interaction between N-WASP and the Arp2/3 Complex Links Cdc42-Dependent Signals to Actin Assembly , 1999, Cell.

[28]  G. Schatten,et al.  Profilin and actin-related proteins regulate microfilament dynamics during early mammalian embryogenesis. , 2006, Human reproduction.

[29]  I. Cameron,et al.  Specific PKC isoforms regulate blastocoel formation during mouse preimplantation development. , 2004, Developmental biology.

[30]  Thomas D Pollard,et al.  Regulation of actin filament assembly by Arp2/3 complex and formins. , 2007, Annual review of biophysics and biomolecular structure.

[31]  Matthew D. Welch,et al.  The ARP2/3 complex: an actin nucleator comes of age , 2006, Nature Reviews Molecular Cell Biology.

[32]  A. DePace,et al.  The Human Arp2/3 Complex Is Composed of Evolutionarily Conserved Subunits and Is Localized to Cellular Regions of Dynamic Actin Filament Assembly , 1997, The Journal of cell biology.

[33]  V. Alarcon Cell Polarity Regulator PARD6B Is Essential for Trophectoderm Formation in the Preimplantation Mouse Embryo1 , 2010, Biology of reproduction.

[34]  Niels Volkmann,et al.  The structural basis of actin filament branching by the Arp2/3 complex , 2008, The Journal of cell biology.