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Ect2, an Ortholog of Drosophila’s Pebble, Negatively Regulates Neurite Outgrowth in Neuroblastoma × Glioma Hybrid NG108-15 Cells

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Abstract

To identify genes required for brain development, we previously performed in vivo RNA interference (RNAi) screening in Drosophila embryos. We identified pebble as a gene that disrupts development of the Drosophila nervous system. Although pebble has been shown to be involved in neuronal development of Drosophila in several screens, the involvement of Ect2, a mammalian ortholog of pebble, in mammalian neuronal development has not been addressed. To examine the role of Ect2 in neuronal differentiation, we performed Ect2 RNAi in the mouse neuroblastoma × rat glioma NG108-15 cell line. Depletion of Ect2 resulted in an increased proportion of binucleate cells and morphological differentiation of NG108-15 cells characterized by the outgrowth of neurites. These morphological changes were correlated with an increased level of acetylcholine esterase mRNA. In addition, expression of Ect2 was decreased in differentiated NG108-15 cells induced by dibutyryl cyclic AMP. These findings indicate that Ect2 negatively regulates the differentiation of NG108-15 cells and suggest that Ect2 may play a role in neuronal differentiation and brain development in vivo.

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References

  • Amano T, Richelson E, Nirenberg M (1972) Neurotransmitter synthesis by neuroblastoma clones (neuroblast differentiation-cell culture-choline acetyltransferase-acetylcholinesterase-tyrosine hydroxylase-axons-dendrites). Proc Natl Acad Sci USA 69:258–263

    Article  PubMed  CAS  Google Scholar 

  • Durand B, Raff M (2000) A cell-intrinsic timer that operates during oligodendrocyte development. Bioessays 22:64–71

    Article  PubMed  CAS  Google Scholar 

  • Gotz M, Huttner WB (2005) The cell biology of neurogenesis. Nat Rev Mol Cell Biol 6(10):777–788

    Article  PubMed  Google Scholar 

  • Govek EE, Newey SE, Van Aelst L (2005) The role of the Rho GTPases in neuronal development. Genes Dev 19:1–49

    Article  PubMed  CAS  Google Scholar 

  • Hamprecht B (1977) Structural, electrophysiological, biochemical, and pharmacological properties of neuroblastoma-glioma cell hybrids in cell culture. Int Rev Cytol 49:99–170

    Article  PubMed  CAS  Google Scholar 

  • Higashida H, Kato T, Kano-Tanaka K, Okuya M, Miyake A, Tanaka T (1981) Proliferation and synapse formation of neuroblastoma glioma hybrid cells: effects of glia maturation factor. Brain Res 214:287–299

    Article  PubMed  CAS  Google Scholar 

  • Higashida H, Streaty RA, Klee W, Nirenberg M (1986) Bradykinin-activated transmembrane signals are coupled via No or Ni to production of inositol 1,4,5-trisphosphate, a second messenger in NG108-15 neuroblastoma-glioma hybrid cells. Proc Natl Acad Sci USA 83:942–946

    Article  PubMed  CAS  Google Scholar 

  • Islam MS, Tsuji T, Higashida C, Takahashi M, Higashida H, Koizumi K (2010) Expression of a Rho guanine nucleotide exchange factor, Ect2, in the developing mouse pituitary. J Neuroendocrinol 22:477–482

    Article  PubMed  CAS  Google Scholar 

  • Ivanov AI, Rovescalli AC, Pozzi P, Yoo S, Mozer B, Li HP, Yu SH, Higashida H, Guo V, Spencer M, Nirenberg M (2004) Genes required for Drosophila nervous system development identified by RNA interference. Proc Natl Acad Sci USA 101:16216–16221

    Article  PubMed  CAS  Google Scholar 

  • Kishi K, Sasaki T, Kuroda S, Itoh T, Takai Y (1993) Regulation of cytoplasmic division of Xenopus embryo by rho p21 and its inhibitory GDP/GTP exchange protein (rho GDI). J Cell Biol 120:1187–1195

    Article  PubMed  CAS  Google Scholar 

  • Koizumi K, Higashida H, Yoo S, Islam MS, Ivanov AI, Guo V, Pozzi P, Yu SH, Rovescalli AC, Tang D, Nirenberg M (2007) RNA interference screen to identify genes required for Drosophila embryonic nervous system development. Proc Natl Acad Sci USA 104:5626–5631

    Article  PubMed  CAS  Google Scholar 

  • Kraut R, Menon K, Zinn K (2001) A gain-of-function screen for genes controlling motor axon guidance and synaptogenesis in Drosophila. Curr Biol 11:417–430

    Article  PubMed  CAS  Google Scholar 

  • Mabuchi I, Hamaguchi Y, Fujimoto H, Morii N, Mishima M, Narumiya S (1993) A rho-like protein is involved in the organisation of the contractile ring in dividing sand dollar eggs. Zygote 1:325–331

    Article  PubMed  CAS  Google Scholar 

  • MacDermot J, Higashida H, Wilson SP, Matsuzawa H, Minna J, Nirenberg M (1979) Adenylate cyclase and acetylcholine release regulated by separate serotonin receptors of somatic cell hybrids. Proc Natl Acad Sci USA 76:1135–1139

    Article  PubMed  CAS  Google Scholar 

  • McGee R, Simpson P, Christian C, Mata M, Nelson P, Nirenberg M (1978) Regulation of acetylcholine release from neuroblastoma × glioma hybrid cells. Proc Natl Acad Sci USA 75:1314–1318

    Article  PubMed  CAS  Google Scholar 

  • Miki N, Hayashi Y, Higashida H (1981) Characterization of chick gizzard extract that promotes neurite outgrowth in cultured ciliary neurons. J Neurochem 37:627–633

    Article  PubMed  CAS  Google Scholar 

  • Minna J, Glazer D, Nirenberg M (1972) Genetic dissection of neural properties using somatic cell hybrids. Nat New Biol 235:225–231

    PubMed  CAS  Google Scholar 

  • Nelson P, Christian C, Nirenberg M (1976) Synapse formation between clonal neuroblastoma × glioma hybrid cells and striated muscle cells. Proc Natl Acad Sci USA 73:123–127

    Article  PubMed  CAS  Google Scholar 

  • Nirenberg M, Wilson S, Higashida H, Rotter A, Krueger K, Busis N, Ray R, Kenimer JG, Adler M (1983) Modulation of synapse formation by cyclic adenosine monophosphate. Science 222:794–799

    Article  PubMed  CAS  Google Scholar 

  • Prokopenko SN, Brumby A, O’Keefe L, Prior L, He Y, Saint R, Bellen HJ (1999) A putative exchange factor for Rho1 GTPase is required for initiation of cytokinesis in Drosophila. Genes Dev 13:2301–2314

    Article  PubMed  CAS  Google Scholar 

  • Prokopenko SN, He Y, Lu Y, Bellen HJ (2000) Mutations affecting the development of the peripheral nervous system in Drosophila: a molecular screen for novel proteins. Genetics 156:1691–1715

    PubMed  CAS  Google Scholar 

  • Puro DG, Nirenberg M (1976) On the specificity of synapse formation. Proc Natl Acad Sci USA 73:3544–3548

    Article  PubMed  CAS  Google Scholar 

  • Reiter LT, Seagroves TN, Bowers M, Bier E (2006) Expression of the Rho-GEF Pbl/ECT2 is regulated by the UBE3A E3 ubiquitin ligase. Hum Mol Genet 15:2825–2835

    Article  PubMed  CAS  Google Scholar 

  • Schmidt A, Hall A (2002) Guanine nucleotide exchange factors for Rho GTPases: turning on the switch. Genes Dev 16:1587–1609

    Article  PubMed  CAS  Google Scholar 

  • Scoumanne A, Chen X (2006) The epithelial cell transforming sequence 2, a guanine nucleotide exchange factor for Rho GTPases, is repressed by p53 via protein methyltransferases and is required for G1-S transition. Cancer Res 66:6271–6279

    Article  PubMed  CAS  Google Scholar 

  • Seeds NW, Gilman AG, Amano T, Nirenberg MW (1970) Regulation of axon formation by clonal lines of a neural tumor. Proc Natl Acad Sci USA 66:160–167

    Article  PubMed  CAS  Google Scholar 

  • Smallhorn M, Murray MJ, Saint R (2004) The epithelial-mesenchymal transition of the Drosophila mesoderm requires the Rho GTP exchange factor Pebble. Development 131:2641–2651

    Article  PubMed  CAS  Google Scholar 

  • Takahashi T, Nowakowski RS, Caviness VS Jr (1995) The cell cycle of the pseudostratified ventricular epithelium of the embryonic murine cerebral wall. J Neurosci 15:6046–6057

    PubMed  CAS  Google Scholar 

  • Tatsumoto T, Xie X, Blumenthal R, Okamoto I, Miki T (1999) Human ECT2 is an exchange factor for Rho GTPases, phosphorylated in G2/M phases, and involved in cytokinesis. J Cell Biol 147:921–928

    Article  PubMed  CAS  Google Scholar 

  • Wilson SH, Schrier BK, Farber JL, Thompson EJ, Rosenberg RN, Blume AJ, Nirenberg MW (1972) Markers for gene expression in cultured cells from the nervous system. J Biol Chem 247:3159–3169

    PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Biophysical Genetics, Graduate School of Medical Science, Kanazawa, 920-8640, Japan

    Takahiro Tsuji, Chiharu Higashida, Mohammad Saharul Islam, Keita Koizumi & Haruhiro Higashida

  2. Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency, Tokyo, 102-0075, Japan

    Chiharu Higashida & Haruhiro Higashida

  3. Research Center for Child Mental Development, Kanazawa, 920-8640, Japan

    Keita Koizumi & Haruhiro Higashida

  4. Genome Biotechnology Laboratory, Kanazawa Institute of Technology, Ishikawa, 924-0838, Japan

    Yasumasa Yoshida & Mitsuko Dohmoto

Authors
  1. Takahiro Tsuji
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  2. Chiharu Higashida
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  3. Yasumasa Yoshida
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  4. Mohammad Saharul Islam
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  5. Mitsuko Dohmoto
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  6. Keita Koizumi
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  7. Haruhiro Higashida
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Correspondence to Haruhiro Higashida.

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Takahiro Tsuji and Chiharu Higashida equally contributed to this work.

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Tsuji, T., Higashida, C., Yoshida, Y. et al. Ect2, an Ortholog of Drosophila’s Pebble, Negatively Regulates Neurite Outgrowth in Neuroblastoma × Glioma Hybrid NG108-15 Cells. Cell Mol Neurobiol 31, 663–668 (2011). https://doi.org/10.1007/s10571-011-9668-3

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  • DOI: https://doi.org/10.1007/s10571-011-9668-3

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