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Spatial organization of cytoskeletal actin filaments and their regulatory proteins revealed by advanced methods in electron microscopy.

Research Project

Project/Area Number 22370056
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Biophysics
Research InstitutionNagoya University

Principal Investigator

USUKURA Jiro  名古屋大学, エコトピア科学研究所, 教授 (30143415)

Co-Investigator(Kenkyū-buntansha) SUZUKI Motoshi  名古屋大学, 大学院・医学系研究科, 講師 (80236017)
Project Period (FY) 2010 – 2012
Project Status Completed (Fiscal Year 2012)
Budget Amount *help
¥19,110,000 (Direct Cost: ¥14,700,000、Indirect Cost: ¥4,410,000)
Fiscal Year 2012: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000)
Fiscal Year 2011: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000)
Fiscal Year 2010: ¥8,190,000 (Direct Cost: ¥6,300,000、Indirect Cost: ¥1,890,000)
Keywordsアクチン / 空間構造 / 細胞骨格 / 核膜 / 電子顕微鏡 / 中間径線維 / 微小管 / ストレス線維 / アクチン線維 / 細胞周期 / ネスプリン / 生物物理 / ミオシン / 制御タンパク質 / 発生・分化
Research Abstract

Current study aims to reveal the spatial organization of cytoskeletal actin filaments in the periphery of nuclear envelope. Cytoskeletal actin filaments in living cell have been investigated so far exclusively withfluorescent light microscopy. Therefore, observation was restricted on the stress fiber with relatively strong fluorescence and at ventral side of the cell. In order to detect real spatial structure of cytoskeleton, high voltage TEM (1000 KV), high resolution SEM and immuno-freeze etching technique were applied to unroofed whole cells. Nuclear envelope was supplied with many cytoskeleton including intermediate filaments, actin filaments and microtubules. In particular, intermediate filaments, vimentin, extending from nuclear pore like a rosette formed complicated meshwork and covered the total surface. Many actin filaments were attached to nuclear envelope as well. Since S1 decoration showed both pointed and barbed ends clearly in the surface of nucleus, there seemed to be the terminations and origins of actin filaments on the nuclear envelope. Unfortunately, however, we were not able to trace entire actin filaments on the nuclear envelope, because all actin filaments were associated or covered with vimentin filaments on the surface of nucleus. Under careful observation of many electron micrographs, many actin filaments appeared to extend from nuclear pores. Indeed, Nesprin 1, actin binding protein, was detected inthe periphery of nuclear pores. Actin filaments extending from nuclear envelope were bundled gradually to form stress fibers. These are completely new concept on nuclear related actin filaments.

Report

(4 results)
  • 2012 Annual Research Report   Final Research Report ( PDF )
  • 2011 Annual Research Report
  • 2010 Annual Research Report
  • Research Products

    (42 results)

All 2013 2012 2011 2010 Other

All Journal Article (13 results) (of which Peer Reviewed: 13 results) Presentation (26 results) (of which Invited: 2 results) Remarks (3 results)

  • [Journal Article] Development of an environmental high-voltage electron microscope for reaction science2013

    • Author(s)
      Tanaka N, Usukura J, Kusunoki M, Saito Y, Sasaki K, Tanji T, Muto S, Arai S
    • Journal Title

      Microscopy

      Volume: 62 Pages: 205-215

    • NAID

      120005510922

    • Related Report
      2012 Final Research Report
    • Peer Reviewed
  • [Journal Article] Development of an Environmental High-Voltage Electron Microscope for Reaction Science2013

    • Author(s)
      N. Tanaka
    • Journal Title

      Microscopy

      Volume: 62 Issue: 1 Pages: 205-215

    • DOI

      10.1093/jmicro/dfs095

    • Related Report
      2012 Annual Research Report
    • Peer Reviewed
  • [Journal Article] MYBPH inhibits NM IIA assembly via direct interaction with NMHC IIA and reduces cell motility2012

    • Author(s)
      Hosono Y, Usukura J, Yamaguchi T. Yanagisawa K, Suzuki M, Takahashi T
    • Journal Title

      Biochem. Biophys. Res. Commun.

      Volume: 428 Pages: 173178-173178

    • Related Report
      2012 Final Research Report
    • Peer Reviewed
  • [Journal Article] Use of the unrooifng technique for atomic force microscopic imaging of the intra-cellular cytoskeleton under aqueous conditions.2012

    • Author(s)
      Usukura J, Yoshimura A, Minakata S, Youn D, Ahn J, Cho SJ
    • Journal Title

      J. Electron Microsc

      Volume: 61 Pages: 321-326

    • Related Report
      2012 Final Research Report
    • Peer Reviewed
  • [Journal Article] MYBPH inhibits NM IIA assembly via direct interaction with NMHC IIA and reduces cell motility.2012

    • Author(s)
      Hosono Y
    • Journal Title

      Biochem. Biophys. Res. Commun.

      Volume: 428 Issue: 1 Pages: 173-178

    • DOI

      10.1016/j.bbrc.2012.10.036

    • Related Report
      2012 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Use of the unrooifng technique for atomic force microscopic imaging of the intra-cellular cytoskeleton under aqueous conditions.2012

    • Author(s)
      Usukura J
    • Journal Title

      J. Electron Microsc.

      Volume: 61 Issue: 5 Pages: 321-326

    • DOI

      10.1093/jmicro/dfs055

    • Related Report
      2012 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Low glucose under hypoxic conditions induces unfolded protein response and produces reactive oxygen species in lens epithelial cells.2012

    • Author(s)
      Elanchezhian R
    • Journal Title

      Cell Death Dis.

      Volume: 3 Issue: 4 Pages: e301-e301

    • DOI

      10.1038/cddis.2012.40

    • Related Report
      2012 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Hybrid liposomes affect cellular lipid constituents and caveolae structure2012

    • Author(s)
      Cao K, Tanaka K, Komizu Y, Tamiya-Koizumi K, Murate T, Ueoka R, Kyogashima M, Usukura J, Takahashi T, Suzuki M
    • Journal Title

      Bioorg Med Chem Lett

      Volume: 22 Pages: 1731-1733

    • Related Report
      2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Spindle microtubules generate tension-dependent changes in the distribution of inner kinetochore proteins.2011

    • Author(s)
      Suzuki A, Hori T, Nishino T, Usukura J, Miyagi A, Morikawa K, Fukagawa T
    • Journal Title

      J. Cell Biol

      Volume: 193 Pages: 125-140

    • Related Report
      2012 Final Research Report
    • Peer Reviewed
  • [Journal Article] Spindle microtubules generate tension-dependent changes in the distribution of inner kinetochore2011

    • Author(s)
      Suzuki A, Hori T, Nishino T, Usukura J, Miyagi A, Morikawa K, Fukagawa T
    • Journal Title

      J Cell Biol

      Volume: 193 Pages: 125-140

    • Related Report
      2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Spindle microtubules generate tension-dependent changes in the distribution ofinner kinetochore proteins2011

    • Author(s)
      Suzuki A, Hori T, Nishino T, Usukura J, Miyagi A, Morikawa K, Fukagawa T
    • Journal Title

      J.Cell Biol.

      Volume: 193 Pages: 125-140

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Mechanism for the regulation of mammalian cGMP phosphodiesterase6.2 : isolation and characterization of the transducin-activated form.2010

    • Author(s)
      Yamazaki A, Tatsumi M, Bondarenko VA, Kurono S, Komori N, Matsumoto H, Matsuura I, Hayashi F, Yamazaki RK, Usukura J.
    • Journal Title

      Mol Cell Biochem.

      Volume: 339 Pages: 235-251

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Mechanism for the regulation of mammalian cGMP phosphodiesterase6.1 : identification of its history subunit complexes and their roles.2010

    • Author(s)
      Yamazaki A, Bondarenko VA, Matsuura I, Tatsumi M, Kurono S, Komori N, Matsumoto H, Hayashi F, Yamazaki RK, Usukura J.
    • Journal Title

      Mol Cell Biochem.

      Volume: 339 Pages: 215-233

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Presentation] Spatial structure of peri-unclear actin cytoskeleton2013

    • Author(s)
      Usukura J, Minakata S
    • Organizer
      The 14thInternational Membrane Research Forum
    • Place of Presentation
      Kyoto, Japan
    • Related Report
      2012 Final Research Report
  • [Presentation] Spatial structure of peri-unclear actin cytoskeleton2013

    • Author(s)
      Usukura J
    • Organizer
      The 14th International Membrane Research Forum,
    • Place of Presentation
      Kyoto Japan
    • Related Report
      2012 Annual Research Report
    • Invited
  • [Presentation] アクチン細胞骨格の空間構造2012

    • Author(s)
      臼倉治郎、南方志帆
    • Organizer
      第117回日本解剖学会
    • Place of Presentation
      山梨大学甲府キャンパス(招待講演)
    • Year and Date
      2012-03-28
    • Related Report
      2011 Annual Research Report
  • [Presentation] Spatial organization of actin and intermediate filaments on the nuclear envelope2012

    • Author(s)
      Usukura J, Minakata S
    • Organizer
      The American Society for Cell Biology
    • Place of Presentation
      Moscone Center San Francisco, CA, USA
    • Related Report
      2012 Final Research Report
  • [Presentation] Spatial structure of actin cytoskeletons around the nuclear envelope2012

    • Author(s)
      Usukura J, Minakata S
    • Organizer
      The 4thEMBO meeting
    • Place of Presentation
      Nice Acropolice France
    • Related Report
      2012 Final Research Report
  • [Presentation] Spatial structure of cytoplasmic actin filaments with special reference to the association with nuclear envelope2012

    • Author(s)
      Usukura J, Minakata S
    • Organizer
      Microscopy & Microanalysis
    • Place of Presentation
      Phoenix Convention Center Arizona USA
    • Related Report
      2012 Final Research Report
  • [Presentation] Spatial organization of actin and intermediate filaments on the nuclear envelope2012

    • Author(s)
      Usukura J
    • Organizer
      The American Society for Cell Biology
    • Place of Presentation
      San Francisco CA, USA
    • Related Report
      2012 Annual Research Report
  • [Presentation] Spatial structure of actin cytoskeletons around the nuclear envelope2012

    • Author(s)
      Usukura J
    • Organizer
      The 4th EMBO meeting
    • Place of Presentation
      NIce, France
    • Related Report
      2012 Annual Research Report
  • [Presentation] Spatial structure of cytoplasmic actin filaments with special reference to the association with nuclear envelope2012

    • Author(s)
      Usukura J
    • Organizer
      Microscopy & Microanalysis 2012
    • Place of Presentation
      Phoenix, AZ, USA
    • Related Report
      2012 Annual Research Report
    • Invited
  • [Presentation] Spatial structure of actin cytoskeletons associated with nuclear membrane2011

    • Author(s)
      J.Usukura
    • Organizer
      The 7^<th> International Forum on Oxidative Stress and Aging
    • Place of Presentation
      Medical University of Gdansk, Poland(招待講演)
    • Year and Date
      2011-11-23
    • Related Report
      2011 Annual Research Report
  • [Presentation] Spatial structure and specificity of cytoskeletal actin filaments in pigment epithelial cells2011

    • Author(s)
      J.Usukura
    • Organizer
      ARVO International Society for Ocular Cell Biology Conference
    • Place of Presentation
      Vancouver Marriott Pinnacle Downtown Hotel(招待講演)
    • Year and Date
      2011-09-08
    • Related Report
      2011 Annual Research Report
  • [Presentation] Spatial structure of actin cytoskeletons associated with nuclear membrane2011

    • Author(s)
      Usukura J, Minakata S
    • Organizer
      The American Society for Cell Biology
    • Place of Presentation
      Colorado Convention Center Denver, USA
    • Related Report
      2012 Final Research Report
  • [Presentation] Spatial structure and specificity of cytoskeletal actin filaments in pigment epithelial cells ARVO International2011

    • Author(s)
      Usukura J
    • Organizer
      Society for Ocular Cell Biology Conference
    • Place of Presentation
      Vancouver Marriott Pinnacle Downtown Hotel, Vancouver Canada (invited lecture)
    • Related Report
      2012 Final Research Report
  • [Presentation] Spatial structure of actin cytoskeletons associated with nuclear membrane2011

    • Author(s)
      J.Usukura, S.Minakata
    • Organizer
      2011 annual meeting of the American Society for Cell Biology
    • Place of Presentation
      Denver convention center
    • Related Report
      2011 Annual Research Report
  • [Presentation] Spatial structure of cytoskeleton in whole mount cell2011

    • Author(s)
      Minakata S, Usukura J
    • Organizer
      Eleventh International Symposium on Biomimetic Materials Processing (BMMP-11)
    • Place of Presentation
      Noyori Conference Hall, Nagoya Japan
    • Related Report
      2010 Annual Research Report
  • [Presentation] Spatial arrangement of cytoskeletal actin filaments with reference to the association with nuclear membrane2011

    • Author(s)
      Usukura J, Minakata S
    • Organizer
      Eleventh International Symposium on Biomimetic Materials Processing (BMMP-11)
    • Place of Presentation
      Noyori Conference Hall, Nagoya Japan
    • Related Report
      2010 Annual Research Report
  • [Presentation] 細胞質細胞骨格の空間構造解析2011

    • Author(s)
      南方志帆, 臼倉治郎
    • Organizer
      生体運動研究合同班会議
    • Place of Presentation
      大阪市立大学
    • Related Report
      2010 Annual Research Report
  • [Presentation] Spatial Arrangement of Cytoplasmic Cytoskeletons Associated with Nuclear Membrane and its Changes during cell cycle2010

    • Author(s)
      Usukura J, Minakata S
    • Organizer
      The American Society for Cell Biology 50^<th> Annual Meeting
    • Place of Presentation
      Pennsylvania Convention Center, PA, USA
    • Related Report
      2010 Annual Research Report
  • [Presentation] The spatial structure of the actin filament in live cells2010

    • Author(s)
      Usukura J
    • Organizer
      GCOE/Structural Biology Research Center International Symposium
    • Place of Presentation
      Noyori Conference Hall Nagoya, Japan
    • Related Report
      2010 Annual Research Report
  • [Presentation] Spatial structure of cytoskeleton in whole mount cell2010

    • Author(s)
      Minakata S, Usukura J
    • Organizer
      The 4^<th> Mechanobiology Workshop and Biophysical Society Joint Meeting
    • Place of Presentation
      National University of Singapore, Singapore
    • Related Report
      2010 Annual Research Report
  • [Presentation] Spatial Arrangement of Cytoskeletal Actin filaments and spatial specificity of actin binding proteins2010

    • Author(s)
      Usukura J, Yoshimura A, Minakata S
    • Organizer
      The 4^<th> Mechanobiology Workshop and Biophysical Society Joint Meeting
    • Place of Presentation
      National University of Singapore, Singapore
    • Related Report
      2010 Annual Research Report
  • [Presentation] Spatial structure of cytoskeleton in whole mount cell and its changes during cell cycle revealed by high voltage STEM(1000KV)2010

    • Author(s)
      Usukura J, Minakata S, Hirashima R, Arai S, Tanaka N
    • Organizer
      The 17^<th> IFSM International Microscopy Congress
    • Place of Presentation
      Rio de Janeiro, Brazil
    • Related Report
      2010 Annual Research Report
  • [Presentation] Development of high-voltage electron microscope for reaction science2010

    • Author(s)
      Tanaka N, Usukura J, Kusnoki M, Kuroda K, Saito Y, Tanji T, Muto S, Arai
    • Organizer
      The 17^<th> IFSM International Microscopy Congress
    • Place of Presentation
      Rio de Janeiro, Brazil
    • Related Report
      2010 Annual Research Report
  • [Presentation] 細胞質内の細胞骨格の空間構造2010

    • Author(s)
      平嶋涼子, 臼倉治郎
    • Organizer
      日本顕微鏡学会第66回学術講演会
    • Place of Presentation
      名古屋国際会議場
    • Related Report
      2010 Annual Research Report
  • [Presentation] 全截標本を用いた超高圧電子顕微鏡による細胞質細胞骨格の空間構造解析2010

    • Author(s)
      臼倉治郎, 平嶋涼子, 南方志帆, 荒井重勇
    • Organizer
      日本顕微鏡学会第66回学術講演会
    • Place of Presentation
      名古屋国際会議場
    • Related Report
      2010 Annual Research Report
  • [Presentation] 反応科学超高圧電子顕微鏡の開発2010

    • Author(s)
      田中信夫, 臼倉治郎, 楠美智子, 黒田光太郎, 斉藤弥八, 丹司敬義, 武藤俊介, 荒井重勇
    • Organizer
      日本顕微鏡学会第66回学術講演会
    • Place of Presentation
      名古屋国際会議場
    • Related Report
      2010 Annual Research Report
  • [Remarks]

    • URL

      http://www.numse.nagoya-u.ac.jp/bio/

    • Related Report
      2012 Final Research Report
  • [Remarks] 臼倉研究室

    • URL

      http://www.numse.nagoya-u.ac.jp/bio/

    • Related Report
      2012 Annual Research Report
  • [Remarks]

    • URL

      http://www.numse.nagoya-u.ac.jp/bio

    • Related Report
      2010 Annual Research Report

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Published: 2010-08-23   Modified: 2019-07-29  

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