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Transgenic Mouse Line with Green-fluorescent Protein-labeled Centrin 2 allows Visualization of the Centrosome in Living Cells

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Abstract

The centrosome plays diverse roles throughout the cellular mitotic cycle and in post-mitotic cells. Analysis of centrosome position and dynamics in living murine cells has been limited due to a lack of adequate reporters and currently requires either cell fixation/immunostaining or transfection with centrosome reporters. Here we describe the generation and characterization of a transgenic mouse line that constitutively expresses green fluorescent protein-labeled Centrin-2 (GFP-CETN2). The phenotype of the mouse is indistinguishable from wild-type and it displays a single pair of fluorescent centrioles in cells of every organ and time point examined. This model will be helpful for visualizing the centrosome in multiple experimental conditions.

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References

  • Araki M, Masutani C, Takemura M, Uchida A, Sugasawa K, Kondoh J, Ohkuma Y and Hanaoka F (2001) Centrosome protein centrin 2/caltractin 1 is part of the xeroderma pigmentosum group C complex that initiates global genome nucleotide excision repair. J Biol Chem 276: 18665–18672.

    CAS  PubMed  Google Scholar 

  • Baron AT, Greenwood TM, Bazinet CW and Salisbury JL (1992) Centrin is a component of the pericentriolar lattice. Biol Cell 76: 383–388.

    Article  CAS  PubMed  Google Scholar 

  • Beisson J and Wright M (2003) Basal body/centriole assembly and continuity. Curr Opin Cell Biol 15: 96–104.

    Article  CAS  PubMed  Google Scholar 

  • Bobinnec Y, Khodjakov A, Mir LM, Rieder CL, Edde B and Bornens M (1998) Centriole disassembly in vivo and its effect on centrosome structure and function in vertebrate cells. J Cell Biol 143: 1575–1589.

    Article  CAS  PubMed  Google Scholar 

  • Chenn A, Zhang YA, Chang BT and McConnell SK (1998) Intrinsic polarity of mammalian neuroepithelial cells. Mol Cell Neurosci 11: 183–193.

    Article  CAS  PubMed  Google Scholar 

  • Connolly JA, Kiosses BW and Kalnins VI (1986) Centrioles are lost as embryonic myoblasts fuse into myotubes in vitro. Eur J Cell Biol 39: 341–345.

    CAS  PubMed  Google Scholar 

  • Danowski BA, Khodjakov A and Wadsworth P (2001) Centrosome behavior in motile HGF-treated PtK(2) cells expressing GFP-gamma tubulin. Cell Motil Cytos 50: 59–68.

    CAS  Google Scholar 

  • D'Assoro AB, Stivala F, Barrett S, Ferrigno G and Salisbury JL (2001) GFP-centrin as a marker for centriole dynamics in the human breast cancer cell line MCF-7. Ital J Anat Embryol 106: 103–110.

    PubMed  Google Scholar 

  • Debec A, Detraves C, Montmory C, Geraud G and Wright M (1995) Polar organization of gamma-tubulin in acentriolar mitotic spindles of Drosophila melanogaster cells. J Cell Sci 108 (Pt 7): 2645–2653.

    CAS  PubMed  Google Scholar 

  • Deng CX (2002) Roles of BRCA1 in centrosome duplication. Oncogene 21: 6222–6227.

    CAS  PubMed  Google Scholar 

  • de Saint Phalle B and Sullivan W (1998) Spindle assembly and mitosis without centrosomes in parthenogenetic Sciara embryos. J Cell Biol 141: 1383–1391.

    Article  CAS  PubMed  Google Scholar 

  • Dictenberg JB, Zimmerman W, Sparks CA, Young A, Vidair C, Zheng Y et al. (1998) Pericentrin and gamma-tubulin form a protein complex and are organized into a novel lattice at the centrosome. J Cell Biol 141: 163–174.

    Article  CAS  PubMed  Google Scholar 

  • Diviani D and Scott JD (2001) AKAP signaling complexes at the cytoskeleton. J Cell Sci 114: 1431–1437.

    CAS  PubMed  Google Scholar 

  • Doxsey S (2001) Re-evaluating centrosome function. Nat Rev Mol Cell Biol 2: 688–698.

    Article  CAS  PubMed  Google Scholar 

  • Doxsey SJ, Stein P, Evans L, Calarco PD and Kirschner M (1994) Pericentrin, a highly conserved centrosome protein involved in microtubule organization. Cell 76: 639–650.

    Article  CAS  PubMed  Google Scholar 

  • Errabolu R, Sanders MA and Salisbury JL (1994) Cloning of a cDNA encoding human centrin, an EF-hand protein of centrosomes and mitotic spindle poles. J Cell Sci 107 (Pt 1): 9–16.

    CAS  PubMed  Google Scholar 

  • Glantz SA (1996) Primer of Biostatistics. 4th edn, McGraw-Hill, New York.

    Google Scholar 

  • Hacker G (2000) The morphology of apoptosis. Cell Tissue Res 301: 5–17.

    CAS  PubMed  Google Scholar 

  • Hart PE, Poynter GM, Whitehead CM, Orth JD, Glantz JN, Busby RC, Barrett SL and Salisbury JL (2001) Characterization of the X-linked murine centrin Cetn2 gene. Gene 264: 205–213.

    Article  CAS  PubMed  Google Scholar 

  • Hinchcliffe EH, Miller FJ, Cham M, Khodjakov A and Sluder G (2001) Requirement of a centrosomal activity for cell cycle progression through G1 into S phase. Science 291: 1547–1550.

    Article  CAS  PubMed  Google Scholar 

  • Hinds JW and Ruffett TL (1971) Cell proliferation in the neural tube: an electron microscopic and golgi analysis in the mouse cerebral vesicle. Z Zellforsch Mikrosk Anat 115: 226–264.

    Article  CAS  PubMed  Google Scholar 

  • Klotz C, Garreau de Loubresse N, Ruiz F and Beisson J (1997) Genetic evidence for a role of centrin-associated proteins in the organization and dynamics of the infraciliary lattice in Paramecium.Cell Motil Cytos 38: 172–186.

    CAS  Google Scholar 

  • Laoukili J, Perret E, Middendorp S, Houcine O, Guennou C, Marano F, Bornens M and Tournier F (2000) Differential expression and cellular distribution of centrin isoforms during human ciliated cell differentiation in vitro. J Cell Sci 113 (Pt 8): 1355–1364.

    CAS  PubMed  Google Scholar 

  • Marshall WF, Vucica Y and Rosenbaum JL (2001) Kinetics and regulation of de novo centriole assembly. Implications for the mechanism of centriole duplication. Curr Biol 11: 308–317.

    CAS  PubMed  Google Scholar 

  • Meads T and Schroer TA (1995) Polarity and nucleation of microtubules in polarized epithelial cells. Cell Motil Cytos 32: 273–288.

    CAS  Google Scholar 

  • Meraldi P, Lukas J, Fry AM, Bartek J and Nigg EA (1999) Centrosome duplication in mammalian somatic cells requires E2F and Cdk2-cyclin A. Nat Cell Biol 1: 88–93.

    CAS  PubMed  Google Scholar 

  • Mussman JG, Horn HF, Carroll PE, Okuda M, Tarapore P, Donehower LA and Fukasawa K (2000) Synergistic induction of centrosome hyperamplification by loss of p53 and cyclin E overexpression. Oncogene 19: 1635–1646.

    Article  CAS  PubMed  Google Scholar 

  • Okabe M, Ikawa M, Kominami K, Nakanishi T and Nishimune Y (1997) 'Green mice' as a source of ubiquitous green cells. FEBS Lett 407: 313–319.

    Article  CAS  PubMed  Google Scholar 

  • Paoletti A, Moudjou M, Paintrand M, Salisbury JL and Bornens M (1996) Most of centrin in animal cells is not centrosomeassociated and centrosomal centrin is confined to the distal lumen of centrioles. J Cell Sci 109 (Pt 13): 3089–3102.

    CAS  PubMed  Google Scholar 

  • Piel M, Meyer P, Khodjakov A, Rieder CL and Bornens M (2000) The respective contributions of the mother and daughter centrioles to centrosome activity and behavior in vertebrate cells.J Cell Biol 149: 317–330.

    Article  CAS  PubMed  Google Scholar 

  • Piel M, Nordberg J, Euteneuer U and Bornens M (2001) Centrosome-dependent exit of cytokinesis in animal cells.Science 291: 1550–1553.

    Article  CAS  PubMed  Google Scholar 

  • Ralston E (1993) Changes in architecture of the Golgi complex and other subcellular organelles during myogenesis. J Cell Biol 120: 399–409.

    Article  CAS  PubMed  Google Scholar 

  • Salisbury JL, Suino KM, Busby R and Springett M(2002) Centrin-2 is required for centriole duplication in mammalian cells. Curr Biol 12: 1287–1292.

    Article  CAS  PubMed  Google Scholar 

  • Schliwa M, Pryzwansky KB and Euteneuer U (1982) Centrosome splitting in neutrophils: an unusual phenomenon related to cell activation and motility. Cell 31: 705–717.

    Article  CAS  PubMed  Google Scholar 

  • Sluder G, Miller FJ and Rieder CL (1989) Reproductive capacity of sea urchin centrosomes without centrioles. Cell Motil Cytos 13: 264–273.

    CAS  Google Scholar 

  • Stearns T, Evans L and Kirschner M (1991) Gamma-tubulin is a highly conserved component of the centrosome. Cell 65: 825–836.

    Article  CAS  PubMed  Google Scholar 

  • Stevenson VA, Kramer J, Kuhn J and Theurkauf WE (2001) Centrosomes and the Scrambled protein coordinate microtubuleindependent actin reorganization. Nat Cell Biol 3: 68–75.

    CAS  PubMed  Google Scholar 

  • Tassin AM, Maro B and Bornens M (1985a) Fate of microtubuleorganizing centers during myogenesis in vitro. J Cell Biol 100: 35–46.

    Article  CAS  PubMed  Google Scholar 

  • Tassin AM, Paintrand M, Berger EG and Bornens M (1985b) The Golgi apparatus remains associated with microtubule organizing centers during myogenesis. J Cell Biol 101: 630–638.

    Article  CAS  PubMed  Google Scholar 

  • White RA, Pan Z and Salisbury JL (2000) GFP-centrin as a marker for centriole dynamics in living cells. Microsc Res Tech 49: 451–457.

    Article  CAS  PubMed  Google Scholar 

  • Wolfrum U, Giessl A and Pulvermuller A (2002) Centrins, a novel group of Ca2+-binding proteins in vertebrate photoreceptor cells. Adv Exp Med Biol 514: 155–178.

    CAS  PubMed  Google Scholar 

  • Xu X, Weaver Z, Linke SP, Li C, Gotay J, Wang XW, Harris CC, Ried T and Deng CX (1999) Centrosome amplification and a defective G2-M cell cycle checkpoint induce genetic instability in BRCA1 exon 11 isoform-deficient cells. Mol Cell 3: 389–395.

    CAS  PubMed  Google Scholar 

  • Yvon AM, Walker JW, Danowski B, Fagerstrom C, Khodjakov A and Wadsworth P (2002) Centrosome reorientation in woundedge cells is cell type specific. Mol Biol Cell 13: 1871–1880.

    Article  CAS  PubMed  Google Scholar 

  • Zheng Y, Jung MK and Oakley BR (1991) Gamma-tubulin is present in Drosophila melanogaster and Homo sapiens and is associated with the centrosome. Cell 65: 817–823.

    Article  CAS  PubMed  Google Scholar 

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Authors and Affiliations

  1. Neurogenetics Laboratory, Department of Neurosciences, University of California, San Diego, USA

    Holden Higginbotham, Stephanie Bielas, Teruyuki Tanaka & Joseph G. Gleeson

  2. Biomedical Sciences Graduate Program, Department of Neurosciences, University of California, San Diego, USA

    Holden Higginbotham

  3. Biology Graduate Program, Department of Neurosciences, University of California, San Diego, USA

    Stephanie Bielas

Authors
  1. Holden Higginbotham
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  2. Stephanie Bielas
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  3. Teruyuki Tanaka
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  4. Joseph G. Gleeson
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Correspondence to Joseph G. Gleeson.

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Higginbotham, H., Bielas, S., Tanaka, T. et al. Transgenic Mouse Line with Green-fluorescent Protein-labeled Centrin 2 allows Visualization of the Centrosome in Living Cells. Transgenic Res 13, 155–164 (2004). https://doi.org/10.1023/B:TRAG.0000026071.41735.8e

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  • DOI: https://doi.org/10.1023/B:TRAG.0000026071.41735.8e

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