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2006 Fiscal Year Final Research Report Summary

Molecular Mechanisms of Exocytotic Prpces and Its Spatio-temporal Regulation

Research Project

Project/Area Number 15200030
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section一般
Research Field Neurochemistry/Neuropharmacology
Research InstitutionSophia University

Principal Investigator

KUMAKURA Konosuke  Sophia University, Faculty of Science and Technology, Professor (70129790)

Co-Investigator(Kenkyū-buntansha) SASAKAWA Nobuyuki  Sophia University, Faculty of Science and Technology, Professor (20187107)
MURAYAMA Norie  Showa Pharmaceutical University, Assistant (90219949)
Project Period (FY) 2003 – 2006
KeywordsChromaffin cell / Exocvtosis / Mvosin-ATPase / Vesicle movement / Vesicle recruitment / Spatio-temporal regulation / PKC / Mvosin-V
Research Abstract

The aim of this research project is to elucidate the molecular mechanism of exocytotic process and its spatio-temporal regulation, and we have obtained the following results.
1) Roles of PKC in vesicle recruitment.
The facilitation of the vesicle recruitment from the reserve pool to the releasable pool by protein kinase C (PKC) is mediated predominantly by PKCα. PKCβ also increases the secretory function, but by a distinct mechanism. Vesicle recruitment was increased by PKC through the vesicle movement at the rate of 0.03-0.06 μm/s, and the facilitation was detected as an increase in the frequency of exocytotic events in the sustained response. The facilitation of the vesicle recruitment by PKC is accompanied with a substantial increase in the vesicle numbers beneathe the plasma membrane. The receptor molecule for the activated PKC, RACK1, seemed to have roles as anchoring and signaling molecules in the regulation.
2) Roles of motor proteins in vesicle recruitment.
The important role of actin-myosin interaction in the recruitment of chromaffin granules to the release sites has been suggested. In the present research, it was suggested that myosin-V is specifically involved in the vesicle recruitment, while myosin-I is involved in the general movement of the vesicles. In addition, myosin-ATPase was shown to be essential for the regulation of the vesicle recruitment in chromaffin cells. The inhibition of myosin-ATPase resulted a decrease in the vesicle numbers beneathe the plasma membrane.
3) Regulation of SNARE complex.
Binding of calmodulin to C-teminus of VAMP-2, and binding between SyntaxinlA and CaMkinase II regulate SNARE complex and are essential for exocytosis.

  • Research Products

    (55 results)

All 2007 2006 2005 2004 2003 Other

All Journal Article (10 results) (of which Peer Reviewed: 5 results) Presentation (45 results)

  • [Journal Article] Characterization of exocytotic events from single PC12 cells:Amperometric studies in native PS12h,DA-loaded PC12h and bovine adrenal chromaffin cells2005

    • Author(s)
      N.Sasakawa
    • Journal Title

      Cell Mol.Neurobiol. 25

      Pages: 774-787

    • Description
      「研究成果報告書概要(和文)」より
    • Peer Reviewed
  • [Journal Article] Myosin-Va regulates exocytosis through the submicromolar Ca^<2+>-dependent binding of syntaxin-1A2005

    • Author(s)
      M.Watanabe
    • Journal Title

      Mol.Biol.Cell 16

      Pages: 4519-4530

    • Description
      「研究成果報告書概要(和文)」より
    • Peer Reviewed
  • [Journal Article] Characterization of exocytotic events from single PC12 cells : Amperometric studies in native PS12h, DA-loaded PC12h and bovine adrenal chromaffin cells2005

    • Author(s)
      N., Sasakawa, N., Murayama, K., Kumakura
    • Journal Title

      Cell Mol. Neurobiol 25

      Pages: 774-787

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Myosin-Va regulates exocytosis through the submicromolar Ca^<2+>-dependent binding of syntaxin-1A2005

    • Author(s)
      M., Watanabe, K., Nomura , A., Ohyama, R., Ishikawa, Y., Komiya, K., Hosaka, E., Yamauchi, H., Taniguchi, N., Sasakawa, K., Kumakura, T., Ushiki, O., Sato, M., Ikebe, M., Igarashi
    • Journal Title

      Mol. Biol. Cell 16

      Pages: 4519-4530

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Spatio-temporal regulation of neurotransmitter release by PKC;studies in adrenal chromaffin cells2004

    • Author(s)
      K.Kumakura
    • Journal Title

      Critical Reviews in Neurobiol. 16(1&2)

      Pages: 169-175

    • Description
      「研究成果報告書概要(和文)」より
    • Peer Reviewed
  • [Journal Article] Tetrapeptides on N-and C-terminal regions of mastoparan in hibit catecholamine release from chromaffin cells by blocking the nicotinic acetylcholine receptor.Cell2004

    • Author(s)
      N.Monoi
    • Journal Title

      Cell.Mol.Neurobiol. 24

      Pages: 37-50

    • Description
      「研究成果報告書概要(和文)」より
    • Peer Reviewed
  • [Journal Article] Spatio-temporal regulation of neurotransmitter release by PKC ; studies in adrenal chromaffin cells2004

    • Author(s)
      K, . Kumakura, N., Sasakawa, N., Murayama, M., Ohara-Imaizumi
    • Journal Title

      Critical Reviews in Neurobiol 16

      Pages: 169-175

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Tetrapeptides on N- and C-terminal regions of mastoparan inhibit catecholamine release from chromaffin cells by blocking the nicotinic acetylcholine receptor2004

    • Author(s)
      N., Monoi, M., Usui, F., Fukuhara, S., Takahash, H., Ito, K., Kumakura, H., Kasai
    • Journal Title

      Cell. Mol. Neurobiol 24

      Pages: 37-50

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] ウシ副腎髄質クロマフィン細胞における分泌顆粒動態の4次元解析

    • Author(s)
      林 光紀
    • Journal Title

      上智大学生命科学研究所紀要 (印刷中)

    • Description
      「研究成果報告書概要(和文)」より
    • Peer Reviewed
  • [Journal Article] Four dimentional analysis of the dynamics of the secretory vesicles in bovine adrenal chromaffin cells. Sophia Life Science Bulletin

    • Author(s)
      Mitsunori, Hayashi
    • Journal Title

      (in press)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Presentation] 4-dimentional analysis of exocytotic events and vesicle movements in chromaffin cells by LSM-5Live system2007

    • Author(s)
      Mitsunori Hayashi
    • Organizer
      第80回 日本薬理学会年会
    • Place of Presentation
      名古屋
    • Year and Date
      20070314-16
    • Description
      「研究成果報告書概要(和文)」より
  • [Presentation] Role of myosin-ATPase on exocytosis in chromaffin cells2007

    • Author(s)
      Mizuki Sekiguchi
    • Organizer
      第80回 日本薬理学会年会
    • Place of Presentation
      名古屋
    • Year and Date
      20070314-16
    • Description
      「研究成果報告書概要(和文)」より
  • [Presentation] Characterization of exocytotic events in ventral midbrain dopaminergic neurons2007

    • Author(s)
      Konosuke Kumakura
    • Organizer
      49回 日本神経化学会大会
    • Place of Presentation
      名古屋
    • Year and Date
      20070314-16
    • Description
      「研究成果報告書概要(和文)」より
  • [Presentation] Inhibitory effects of epigallocatechin-3 gallate on the exocytosis in adrenal chromaffin cells2007

    • Author(s)
      Norie Murayama
    • Organizer
      49回 日本神経化学会大会
    • Place of Presentation
      名古屋
    • Year and Date
      20070314-16
    • Description
      「研究成果報告書概要(和文)」より
  • [Presentation] 4-dimentional analysis of exocytotic events and vesicle movements in chromaffin cells by LSM-5Live system.2007

    • Author(s)
      Mitsunori, Hayashi, Mizuki, Sekiguchi, Fuyuka, Matsuda, Nobuyuki, Sasakawa, Konosuke, Kumakura
    • Organizer
      The 80th Annual Meeting of The Japanese Pharmacological Society
    • Place of Presentation
      Nagoya
    • Year and Date
      20070314-16
    • Description
      「研究成果報告書概要(欧文)」より
  • [Presentation] (2007) Role of myosin-ATPase on exocytosis in chromaffin cells2007

    • Author(s)
      Mizuki, Sekiguchi, Mitsunori, Hayashi, Fuyuki, Matsuda, Nobuyuki, Sasakawa, Konosuke, Kumakura
    • Organizer
      The 80th Annual Meeting of The Japanese Pharmacological Society
    • Place of Presentation
      Nagoya
    • Year and Date
      20070314-16
    • Description
      「研究成果報告書概要(欧文)」より
  • [Presentation] Possible involvement of myosin-ATPase in the spatio-temporal regulation of exocytosis in adrenal chromaffin cells2006

    • Author(s)
      Konosuke Kumakura
    • Organizer
      3rd International Symposium on Salivary Glands in Honor of Niels Stensen.
    • Place of Presentation
      岡崎
    • Year and Date
      20061020-24
    • Description
      「研究成果報告書概要(和文)」より
  • [Presentation] Characterization of exocytotic events in ventral midbrain dopaminergic neurons2006

    • Author(s)
      Konosuke, Kumakura, Satoshi, Yamada, Mizuki, Sekiguchi, Norie, Murayama, Nobuyuki, Sasakawa
    • Organizer
      The 49th Annual Meeting of The Japanese Society for Neurochemistry
    • Place of Presentation
      Nagoya
    • Year and Date
      20060914-16
    • Description
      「研究成果報告書概要(欧文)」より
  • [Presentation] Inhibitory effects of epigallocatechin-3 gallate on the exocytosis in adrenal chromaffin cells2006

    • Author(s)
      Norie, Murayama, Atuko, Otuka, Mitunori, Hayashi, Nobuyuki, Sasakawa, Konosuke, Kumakura
    • Organizer
      The 49th Annual Meeting of The Japanese Society for Neurochemistry
    • Place of Presentation
      Nagoya
    • Year and Date
      20060914-16
    • Description
      「研究成果報告書概要(欧文)」より
  • [Presentation] Effects of epigallocatechin-3 gallate,a green tea catechin,on the regulatory mechanisms of exocytosis in adrenal chromaffin cells2006

    • Author(s)
      Norie Murayama
    • Organizer
      第79回 日本薬理学会年会
    • Place of Presentation
      横浜
    • Year and Date
      20060308-10
    • Description
      「研究成果報告書概要(和文)」より
  • [Presentation] Amperometric detection and characterization of exocytotic events in single ventral midbrain dopaminergic neurons2006

    • Author(s)
      Satoshi Yamada
    • Organizer
      第79回 日本薬理学会年会
    • Place of Presentation
      横浜
    • Year and Date
      20060308-10
    • Description
      「研究成果報告書概要(和文)」より
  • [Presentation] Effects of epigallocatechin-3 gallate, a green tea catechin, on the regulatory mechanisms of exocytosis in adrenal chromaffin cells. J2006

    • Author(s)
      Norie, Murayama, Atuko, Otuka, Nobuyuki, Sasakawa, Konosuke, Kumakura
    • Organizer
      The 79th Annual Meeting of The Japanese Pharmacological Society
    • Place of Presentation
      Yokohama
    • Year and Date
      20060308-10
    • Description
      「研究成果報告書概要(欧文)」より
  • [Presentation] Amperometric detection and characterization of exocytotic events in single ventral midbrain dopaminergic neurons2006

    • Author(s)
      Satoshi, Yamada, Mizuki, Sekiguchi, Norie, Murayama, Nobuyuki, Sasakawa, Konosuke, Kumakura
    • Organizer
      The 79th Annual Meeting of The Japanese Pharmacological Society
    • Place of Presentation
      Yokohama
    • Year and Date
      20060308-10
    • Description
      「研究成果報告書概要(欧文)」より
  • [Presentation] Trafficking for exocytosis;Imaging studies in the chromaffin cell.(Invited Lecture)2006

    • Author(s)
      K.Kumakura
    • Organizer
      13th International Symposium on Chromaffin Cell Biology
    • Place of Presentation
      Pucon,Chile
    • Year and Date
      20060107-12
    • Description
      「研究成果報告書概要(和文)」より
  • [Presentation] Trafficking for exocytosis ; Imaging studies in the chromaffin cell2006

    • Author(s)
      K., Kumakura, N., Sasakawa, N., Murayama, M., Murakami, A., Riva
    • Organizer
      13th International Symposium on Chromaffin Cell Biology
    • Place of Presentation
      Pucon, Chile
    • Year and Date
      20060107-12
    • Description
      「研究成果報告書概要(欧文)」より
  • [Presentation] A single exoxcytotic event in digitonin-permeabilized adrenal chromaffin cells2005

    • Author(s)
      Nobuyuki Sasakawa
    • Organizer
      第48回 日本神経化学会大会
    • Place of Presentation
      福岡
    • Year and Date
      20050928-30
    • Description
      「研究成果報告書概要(和文)」より
  • [Presentation] A single exoxcytotic event in digitonin-permeabilized adrenal chromaffin cells2005

    • Author(s)
      Nobuyuki, Sasakawa, Norie, Murayama, Konosuke, Kumakura
    • Organizer
      The 48th Annual Meeting of The Japanese Society for Neurochemistry
    • Place of Presentation
      Fukuoka
    • Year and Date
      20050928-30
    • Description
      「研究成果報告書概要(欧文)」より
  • [Presentation] Comparative view of the plasma membrane-vesicle transfer dynamics2005

    • Author(s)
      Susumu Terakawa
    • Organizer
      第28回 日本神経科学大会
    • Place of Presentation
      横浜
    • Year and Date
      20050726-28
    • Description
      「研究成果報告書概要(和文)」より
  • [Presentation] Characterization of a single exoxcytotic event in digitonin-permeabilized adrenal chromaffin cells2005

    • Author(s)
      Nobuyuki Sasakawa
    • Organizer
      第78回 日本薬理学会年会
    • Place of Presentation
      横浜
    • Year and Date
      20050322-24
    • Description
      「研究成果報告書概要(和文)」より
  • [Presentation] Characterization of a single exoxcytotic event in digitonin-permeabilized adrenal chromaffin cells2005

    • Author(s)
      Nobuyuki, Sasakawa, Norie, Murayama, Konosuke, Kumakura
    • Organizer
      The 78th Annual Meeting of The Japanese Pharmacological Society
    • Place of Presentation
      Yokohama
    • Year and Date
      20050322-24
    • Description
      「研究成果報告書概要(欧文)」より
  • [Presentation] (2003) Regulation of Exocytotic Events in Single Adrenal Chromaffin Cells2004

    • Author(s)
      N., Sasakawa, K, . Kumakura
    • Organizer
      The 46th Annual Meeting of The Japanese Society for Neurochemistry
    • Place of Presentation
      Niigata
    • Year and Date
      20040924-26
    • Description
      「研究成果報告書概要(欧文)」より
  • [Presentation] Alessandro Riva, Mica Ohara-Imaizumi, Masataka Murakami(2003)Three-dimensional Structure of the Interior of Chromaffin Cells and Choromaffin Granule Motion2004

    • Author(s)
      Konosuke, Kumakura
    • Organizer
      The 46th Annual Meeting of The Japanese Society for Neurochemistry
    • Place of Presentation
      Niigata
    • Year and Date
      20040924-26
    • Description
      「研究成果報告書概要(欧文)」より
  • [Presentation] (2003)Amperometric detection and characterization of exocytosis from single PC12h cells2004

    • Author(s)
      Nobuyuki, Sasakawa, Norie, Murayama, Konosuke, Kumakura
    • Organizer
      The 46th Annual Meeting of The Japanese Society for Neurochemistry
    • Place of Presentation
      Niigata
    • Year and Date
      20040924-26
    • Description
      「研究成果報告書概要(欧文)」より
  • [Presentation] A high efficient transfecdtion method for chromaffin cells2004

    • Author(s)
      Norie, Murayama, Nobuyuki, Sasakawa, Natsue, Yoshimura, Konosuke, Kumakura
    • Organizer
      The 46th Annual Meeting of The Japanese Society for Neurochemistry
    • Place of Presentation
      Niigata
    • Year and Date
      20040924-26
    • Description
      「研究成果報告書概要(欧文)」より
  • [Presentation] Tempo-spatial regulation of the vesicle dynamics by PKC in adrenal chromaffin cells2004

    • Author(s)
      Konosuke Kumakura
    • Organizer
      第47回 日本神経化学会大会
    • Place of Presentation
      大阪
    • Year and Date
      20040921-23
    • Description
      「研究成果報告書概要(和文)」より
  • [Presentation] Regulation of exocytotic events by protein kinase C isoform in single adrenal chromaffin cells2004

    • Author(s)
      Nobuyuki Sasakawa
    • Organizer
      第47回 日本神経化学会大会
    • Place of Presentation
      大阪
    • Year and Date
      20040921-23
    • Description
      「研究成果報告書概要(和文)」より
  • [Presentation] Characteristics of the Ca^<2+>-dependent binding between myosin-V and syntaxin-1A2004

    • Author(s)
      Michitoshi Watanabe
    • Organizer
      第47回 日本神経化学会大会
    • Place of Presentation
      大阪
    • Year and Date
      20040921-23
    • Description
      「研究成果報告書概要(和文)」より
  • [Presentation] Tempo-spatial regulation of the vesicle dynamics by PKC in adrenal chromaffin cells2004

    • Author(s)
      Konosuke, Kumakura, Nobuyuki, Sasakawa, Norie, Murayama
    • Organizer
      The 47th Annual Meeting of The Japanese Society for Neurochemistry
    • Place of Presentation
      Osaka
    • Year and Date
      20040921-23
    • Description
      「研究成果報告書概要(欧文)」より
  • [Presentation] Regulation of exocytotic events by protein kinase C isoform in single adrenal chromaffin cells2004

    • Author(s)
      Nobuyuki, Sasakawa, Norie, Murayama, Konosuke, Kumakura
    • Organizer
      The 47th Annual Meeting of The Japanese Society for Neurochemistry
    • Place of Presentation
      Osaka
    • Year and Date
      20040921-23
    • Description
      「研究成果報告書概要(欧文)」より
  • [Presentation] Ryokinlshikawa, Nobuyuki Sasakawa, Konosuke Kumakura, Michiro Igarashi. Characteristics of the Ca^<2+>-dependent binding between myosin-V and syntaxin-1A2004

    • Author(s)
      Michitosh, i Watanabe, Tatsuo, Ushiki
    • Organizer
      The 47th Annual Meeting of The Japanese Society for Neurochemistry
    • Place of Presentation
      Osaka
    • Year and Date
      20040921-23
    • Description
      「研究成果報告書概要(欧文)」より
  • [Presentation] Characteristic regulation of exocytotic eventsnby PKC isoforms in single adrenal chromaffin cells2004

    • Author(s)
      Nobuyuki Sasakawa
    • Organizer
      第77回 日本薬理学会年会
    • Place of Presentation
      大阪
    • Year and Date
      20040308-10
    • Description
      「研究成果報告書概要(和文)」より
  • [Presentation] Effects of Shiga Toxin on Exocytotic Event in Single Adrenal Chromaffin Cells2004

    • Author(s)
      Nobuyuki Sasakawa
    • Organizer
      第76回 日本薬理学会年会
    • Place of Presentation
      福岡
    • Year and Date
      20040308-10
    • Description
      「研究成果報告書概要(和文)」より
  • [Presentation] Characteristic regulation of exocytotic eventsnby PKC isoforms in single adrenal chromaffin cells2004

    • Author(s)
      Nobuyuki, Sasakawa, Norie, Murayama, Konosuke, Kumakura
    • Organizer
      The 77th Annual Meeting of The Japanese Pharmacological Society
    • Place of Presentation
      Osaka
    • Year and Date
      20040308-10
    • Description
      「研究成果報告書概要(欧文)」より
  • [Presentation] Kumakura(2003)Effects of Shiga Toxin on Exocytotic Event in Single Adrenal Chromaffin Cells2004

    • Author(s)
      Nobuyuki, Sasakawa, Kiyotaka, Nishikawa, Norie, Murayama, Yumiko, Satoh, Yasuhiro Natori, Konosuke
    • Organizer
      The 76th Annual Meeting of The Japanese Pharmacological Society
    • Place of Presentation
      Fukuoka
    • Year and Date
      20040308-10
    • Description
      「研究成果報告書概要(欧文)」より
  • [Presentation] Tempo-spatial regulation of exocytotic events by protein kinase C in adrenal chromaffin cells2004

    • Author(s)
      K.Kumakura
    • Organizer
      6th Biennial Meeting of the Asian-Pacific Society for Neurochemistry
    • Place of Presentation
      Hong Kong
    • Year and Date
      20040200
    • Description
      「研究成果報告書概要(和文)」より
  • [Presentation] Tempo-spatial regulation of exocytotic events by protein kinase C in adrenal chromaffin cells2004

    • Author(s)
      K., Kumakura, N., Saakawa, S., Hosaka, M., Hayashi, N., Murayama
    • Organizer
      The 6th Biennial Meeting of the Asian-Pacific Society for Neurochemistry
    • Place of Presentation
      Hong Kong
    • Year and Date
      20040200
    • Description
      「研究成果報告書概要(欧文)」より
  • [Presentation] 単一クロマフィン細胞におけるアンペロメトリー法によるカテコールアミン分泌の測定解析2004

    • Author(s)
      笹川 展幸
    • Organizer
      第8回 活性アミンに関するワークショップ, ラウンド、テープル「活性アミン研究における新しいアプローチとその応用」
    • Place of Presentation
      東京
    • Year and Date
      2004-08-27
    • Description
      「研究成果報告書概要(和文)」より
  • [Presentation] クロマフィン細胞における分泌顆粒局在の時空的調節2004

    • Author(s)
      熊倉 鴻之助
    • Organizer
      第8回 活性アミンに関するワークショップ
    • Place of Presentation
      東京
    • Year and Date
      2004-08-27
    • Description
      「研究成果報告書概要(和文)」より
  • [Presentation] Regulation of Exocytotic Events in Single Adrenal Chromaffin Cells2003

    • Author(s)
      N.Sasakawa
    • Organizer
      第46回 日本神経化学会大会
    • Place of Presentation
      新潟
    • Year and Date
      20030924-26
    • Description
      「研究成果報告書概要(和文)」より
  • [Presentation] Three-dimensional Structure of the Interior of Chromaffin Cells and Choromaffin Granule Motion2003

    • Author(s)
      Konosuke Kumakura
    • Organizer
      第46回 日本神経化学会大会
    • Place of Presentation
      新潟
    • Year and Date
      20030924-26
    • Description
      「研究成果報告書概要(和文)」より
  • [Presentation] Amperometric detection and characterization of exocytosis from single PC12h cells2003

    • Author(s)
      Nobuyuki Sasakawa
    • Organizer
      第46回 日本神経化学会大会
    • Place of Presentation
      新潟
    • Year and Date
      20030924-26
    • Description
      「研究成果報告書概要(和文)」より
  • [Presentation] A high efficient transfecdtion method for chromaffin cells2003

    • Author(s)
      Norie Murayama
    • Organizer
      第46回 日本神経化学会大会
    • Place of Presentation
      新潟
    • Year and Date
      20030924-26
    • Description
      「研究成果報告書概要(和文)」より
  • [Presentation] Three-dimensional Structure of the Interior of Chromaffin Cells and Choromaffin Granule Motion2003

    • Author(s)
      Konosuke Kumakura
    • Organizer
      Biennial Meeting of the International Society for Neurochemistry
    • Place of Presentation
      Hong Kong
    • Year and Date
      20030800
    • Description
      「研究成果報告書概要(和文)」より
  • [Presentation] Comparative view of the plasma membrane-vesicle transfer dynamics

    • Author(s)
      Susumu, Terakawa, Takashi, Sakurai, Konosuke, Kumakura, Takahi, Tsuboi, Seiji, Yamamoto
    • Organizer
      The 28th Annual Meeting of the Japanese Neuroscience Society
    • Place of Presentation
      Yokohama
    • Year and Date
      00000726-28
    • Description
      「研究成果報告書概要(欧文)」より
  • [Presentation] Possible involvement of myosin-ATPase in the spatio-temporal regulation of exocytosis in adrenal chromaffin cells

    • Author(s)
      Konosuke, Kumakura, Mizuki, Sekiguchi, Mitunori, Hayashi, Nobuyuki, Sasakawa
    • Organizer
      3rd International Symposium on Salivary Glands in Honor of Niels Stensen. Expanded Abst, 81P
    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2010-02-04  

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