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Regulation of cytoplasmic streaming by calcium in plant cells

Research Project

Project/Area Number 02454008
Research Category

Grant-in-Aid for General Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field 植物生理学
Research InstitutionHimeji Institute of Technology

Principal Investigator

SHIMMEN Teruo  Himeji Institute of Technology, Faculty of Science, Department of Life Science, Full professor, 理学部, 教授 (80114510)

Co-Investigator(Kenkyū-buntansha) YOKOTA Etsuo  Himeji Institute of Technology, Faculty of Science, Department of Life Science,, 理学部, 助手 (80212299)
Project Period (FY) 1990 – 1992
Project Status Completed (Fiscal Year 1992)
Budget Amount *help
¥6,200,000 (Direct Cost: ¥6,200,000)
Fiscal Year 1992: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1991: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1990: ¥5,100,000 (Direct Cost: ¥5,100,000)
Keywordscytoplasmic streaming / actomyosin / lily / pollen tube / calcium / ATPase / ミオシン / 低温 / 車軸藻 / 低温顕微鏡 / イオン輸送 / 液胞膜除去細胞
Research Abstract

Purification of myosin from lily pollen tube We succeeded in isolating myosin from lily pollen tube. Pollen tube was homogenized in a solution of low ion ic strength and the crude extract was prepared by centrifugation. The surface of cover glass was treated with the crude extract. When actin filament labeled with fluorescent dye was applied together with Mg.ATP, actin filament showed sliding movement at the glass surface(in vitro motility assay), indicating that myosin was extracted in an active from. Myosin was successively purified by co-precipitation with actin filament of chiken breast muscle, hydroxylapatite column and gel filtration. When the final sample was analyzed with SDS-PAGE, myosin was found to be composed of heavy chain of 170 kD. We succeeded in rising antibody against the heavy chain. In immunoblotting of the crude extract of pollen tube using the antibody, only 170 kD component reacted with the antibody.
ATPase of the myosin sample was activated to 60 fold by adding a … More ctin filament purified from chiken breast muscle. In in vitro motility assay using purified myosin, actin filament moved at the speed of 7.7 mum/sec, which is almost the same as the speed of cytoplasmic streaming in pollen tube. This fact indicates that the myosin is involved in generation of the motive force of cytoplasmic streaming. We are now planning a experiment to examine the calcium sensitivity of myosin.
Mechanism of calcium regulation of cytoplasmic streaming in characean cells In Characean cells, cytoplasmic streamingtransiently stops upon generation of an action potential. Cytoplasm ic streaming stops due to increase of free calcium in the cytoplasm. It has been supposed that phosphorylation of myosin is involved in stopping of cytoplasmic streaming. In the present study, we analyzed the process of stoppage of cytoplasmic streaming upon depolarization of the membrane by increasing potassium concentration of the external medium in Nitella axilliformis.
When the extracellular concentration of potassium was increased from 0.1 mM to 50 mM, membrane depolarized and stayed at the depolarized state. By decreasing the potassium concentration, membrane repolarized. In response to depolarization of the membrane, cytoplasmic streaming quickly stopped and the velocity stayed at a very low level. On repolarization of the membrane by decreasing potassium concentration, velocity of cytoplasmic streaming recovered within a few minutes. When calcium or magnesium at 5 mM was added to the potassium medium, cytoplasmic streaming stopped as in the absence of these divalent cation. In this case, however, velocity gradually recovered without repolarization of the membrane. It is suggested that these divalent cations bind to the plasma membrane and inhibit the signal processing from membrane depolarization to stoppage of cytoplasimic streaming.
Analysis of temperature dependency of cytoplasmic streaming in Characeae Using cryomicroscope, dependency of velocity of cytoplsmic streaming was analyzed in internodal cells of Nitellopsis obtusa. In intact cells, two types of response was observed among cells tested. In one group, velocity linearly decreased in the temperature range of 25-5゚C. In another group, velocity linearly decreased in a range of 25-15゚C, but strongly inhibited at the temperature lower than 10゚C. Same experiment was also carried out in tonoplast-free cells, in which EGTA, a calcium chelator, and PIPES, a pH buffer, was introduced. In all tonoplast-free cells tested, velocity linearly decreased in temperature range of 25-5゚C. At 5゚C, velocity in tonoplast-free cells was three times higher than that in intact cells. The present study suggested that temperature dependency of cytoplasmic streaming reflects not only temperature characteristics of actomyosin involved in generation of the motive force but also disturbance of intracellular homeostasis of these ions at low tempeture. Less

Report

(4 results)
  • 1992 Annual Research Report   Final Research Report Summary
  • 1991 Annual Research Report
  • 1990 Annual Research Report
  • Research Products

    (47 results)

All Other

All Publications (47 results)

  • [Publications] K.Kohama,T.Kohno,K.Hayakawa,Y.Lin,T.Shimmen,A.Inoue,R.Ishikawa: "A novel regulatory effect of myosin light chain kinase from smooth muscle on the ATP-dependent interaction between actin and myosin." Biochem.Biopys.Res.Comm.184. 1204-1211 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] T.Shimmen: "The characean cytoskeleton:Dissecting the streaming mechanism." In ¨The Cytoskeleton of the algae¨,CRC Press. Ed.D.Menzel. 297-314 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] T.Shimmen: "Mechanisms of the cytoplasmic streaming and amoeboid movement." In ¨Molecular and Cellular Aspects of Muscle Conraction and Cell Motility¨,Springer-Verlag. Ed.H.Sugi. 12. 172-205 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] T.Kohno,R.Ishikawa,T.Nagata,K.Kohama,T.Shimmen: "Partial purification of myosin from liliy pollen tubes by monitoring with in vito motility assay." Protoplasma. 170. 77-85 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] H.Yamashita,S.Sugiura,T.Serizawa,T.Sugimoto,M.Iizuka,E.Katayama,T.Shimmen: "Sliding velocity of isolated rabbit cardac myosin corrlates with isozyme distribution." Am.J.Physiol.263. H464-H472 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] S.Sugiura,H.Yamashita,T.Serizawa,M.Iizuka,T.Shimmen,T.Sugimoto: "Active movemant of cardiac myosin on Characeae actin cables." Pflugers Arch European Journal of Physiology. 421. 32-36 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] 新免 輝男: "細胞の運動" 裳華房, 141 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] 新免 輝男: "細胞の生物学" 岩波書店, 180 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Kohno, T., S. Chaen, T., Shimmen: "Characterization of the translocater associated with pollen tube organelles." Protoplasma. 154. 179-183 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Oiwa, K., S. Chaen, E. Kamitsubo, T. Shimmen, H. Sugi: "Steady-state force-velocity relation in the ATP-dependent sliding movement of myosin-coated beads on actin cabeles in vitro studied with a centrifuge microscope." Proc. Natl. Acad. Sci. USA. 87. 7893-7897 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Shimmen, T., Y.-L. Xu. T. Kohno: "Inhibition of cytoplasmic streaming by sulfate in characean cells." Protoplasma. 158. 39-44 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Tester, M., T. Shimmen, M. J. Beilby, E.A.C. MacRobbie: The transport of ions across the tonoplast of Charophytes: "A study using permeabilized cell." Membrane Transport in Plants and Fungi. 112-115 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] T. Shimmen, E.A.C. MacRobbie: "Proton pumping at the tonoplast of permeabilized Nitella cells." Membrane Transport in Plants and Fungi. 180-182 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] T. Shimmen: "Green muscle:Regulation of cytoplasmic streaming by action potential" Iden(Japanese). 44. 21-25 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Kohno, T., T. Okagaki, K. Kohama, T. Shimmen: "Pollen extract supports the movement of actin filaments in vitro." Protoplama. 161. 75-77 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Kohama, K., T. Kohno, T. Okagaki, T. Shimmen: "Role of actin in the myosin-linked Ca-regulation of ATP-dependent interaction between actin and myosin of lower eukaryote, Phyosarum polycephalum" Protoplama. 161. 75-77 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] T. Shimmen: "Actin cable as a probe for analysis of myosin movement." Seibutsubutsuri (Japanese). 8-11 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Kohama, K., T. Kohno, K. Hayakawa, Y. Lin, R. Ishikawa, T. Shimmen, A. Inoue: "A novel regulatory effect of myosin light chain kinase from smooth muscle on the ATP-dependent interaction between actin and myosin." Biochem. Biopys. Res. Comm.184. 1204-1211 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Kohno, T., R. Ishikawa, T. Nagata, K. Kohama, T. Shimmen: "Partial purification of myosin from liliy pollen tubes by monitoring with in vito motility assay." Protoplasma. 170. 77-85 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Yamashita, H., S. Sugiura, T. Serizawa, T. Sugimoto M. Iizuka, E. Katayam, T. Shimmen: "Sliding velocity of isolated rabbit cardac myosin corrlates with isozym distribution." Am. J. Physiol. 263. H464-H472 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Mimura, H., T. Shimmen: "Permeabilization of the plasma membrane and maintenance of the integrities of organelles in higher plant cells." Plant & Cell Physiol. 33. 703-707 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Sugiura, S., H. Yamashita, T., Serizawa, M. Iizuka, T. Shimmen. T. Sugimoto: "Active movement of cardiac myosin on Characeae actin cables." Europ. J. Physiol. 421. 32-36 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Shimmen, T.: "The characean cytoskleton: Dissecting the streaming mechanism." The Cytoskeleton of the Algae.297-314 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Shimmen, T.: "Mechanisms of cytoplasmic streaming and amoeboid movement." Advance in Comprative and Environmental Physiology. 12. 172-205 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Shimmen, T.: "Inhibitory regulation of cytoplasmic streaming by Ca^<2+> in plant cells." Calcium Inhibition. 69-90 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Mimura, T., K.-J. Dietz, T. Shimmen: "Regulation of apoplastic ion concentration in barley leaves." Plant Cell Walls as Bioopolymers with Physiol, Functions. 344-346 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Mimura, T., & T. Shimmen: "ATP-dependent carbon transpot in perfused Chara cells." Plant cell Environ.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Shimmen, T., E. Yokota: "Mechanism of cytoplasmic streaming." Idenn(Japanese).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] K.Kohama,T.Kohno,K.Hayakawa,Y.Lin,T.Shimmen,A.Inoue R.Ishikawa: "A novel regulatory effect of myosin light chain kinase from smooth muscle on the ATP-dependent interraction between actin and myosin." Biochem.Biopys.Comm.184. 1204-1211 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] T.Shimmen: "The characean cytoskeleton:Dissecting the streaming mechanism." In “The Cytoskeleton of the algae",CRC Press.ED.D.Menzel. 297-314 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] T.Shimmen: "Mechanisms of the cytoplasmic streaming and amoeboid movement." In “Molecular and Cellular Aspects of Muscle Conraction and Cell Motility",Springer-Verlag.Ed.H.Sugi. 12. 172-205 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] T.Kohno,R.Ishikawa,T.Nagata,K.Kohama,T.Shimmen: "Partial purification of myosin from liliy pollen tubes by monitoring with in vito motility assay." Protoplasma. 170. 77-85 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] H.Yamashita,S.Sugiura,T.Serizawa,T.Sugimoto,M.Iizuka,E.Katayama,T.Shimmen: "Sliding velocity of isolated rabbit cardac myosin corrlates with isozyme distribution." Am.J.Physiol.263. H464-H472 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] S.Sugiura,H.Yamashita,T.Serizawa,M.Iizuka,T.Shimmen,T.Sugimoto: "Active movemant of cardiac myosin on Characeae actin cables." Pflugers Arch European Journal of Physiology.421. 32-36 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 新免 輝男: "細胞の運動" 裳華房, 141 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 新免 輝男: "細胞の生物学" 岩波書店, 180 (1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] Kohno,T.,Okagaki,T.,Kohama.K.and Shimmen,T.: "Pollen extract support the movement of actin filaments in vitro" Protoplasma. 161. 75-77 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Kohamna,K.,Kohno,T.,Okagaki,T.and Shimmen,T.: "Role of actin in the myosinーlinked tegulation of ATPーdependent interaction between actin and myosin of a lower eukaryote,Physarum polycephalum" J.Biochemistry. 110. 508-513 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Yamashita,H.,Sugiura,S.,Serizawa,T.,Sugimoto,T.,Izuka,M.,Katayama,E.and Shimmen,T.: "Sliding velocity of isolalted rabbit cardiac myosin correlates with isozyme distribution" American J.Physiology.

    • Related Report
      1991 Annual Research Report
  • [Publications] Shimmen,T.: "Mechanism of cytoplasmic streaming and amoeboid movement" Advances in Comparative and Environmental Physiology. 12. 172-205 (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] 新免 輝男: "現代植物生理学 4 環境応答" 朝倉書店,

    • Related Report
      1991 Annual Research Report
  • [Publications] 新免 輝男: "緑の筋肉ーシャジクモ類における原形質流動の活動電位による制御" 遺伝. 44. 21-25 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Kohno,T.,Chaen,S.,Shimmen,T.: "Characterization of the translocator associated with pollen tube organelles" Protoplasma. 154. 179-183 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Oiwa,K.,Chaen,S.,Kamitsubo,E.,Shimmen,T.,Sugi,H.: "Steadyーstate forceーvelocity relation in the ATPーdependent sliding movement of myosinーcoated beads on actin cables in vitro studied with a centrifuge microscope" Proc.Natl.Acad.Sci.USA. 87. 7893-7897 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Shimmen,T.,Xu,Y.ーL.,Kohno,T.: "Inhibition of cytoplasmic streaming by sulfate in characean cells." Protoplasma. 158. 39-44 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Kohno,T.,Okagaki,T.,Kohama,K.,Shimmen,T.: "Pollen tube extract supports the movement of actin filaments in vitro." Protoplasma. (1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] 新免 輝男(編,著): "現代植物生理学4巻 環境応答" 朝倉書店, 208 (1991)

    • Related Report
      1990 Annual Research Report

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Published: 1990-04-01   Modified: 2016-04-21  

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