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2007 Fiscal Year Final Research Report Summary

Quantitative Imaging and Simulation Study for Analyzing Exocytosis in PC12 Cell

Research Project

Project/Area Number 18300099
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Bioinformatics/Life informatics
Research InstitutionKeio University

Principal Investigator

OKA Kotaro  Keio University, Faculty of Science and Technology, Professor (10276412)

Co-Investigator(Kenkyū-buntansha) HOTTA Koji  Keio University, Faculty of Science and Thchnology, Research Associate (80407147)
Project Period (FY) 2006 – 2007
KeywordsBiological simulation / Neuroscience / Biophysics / Visualization techniques / Stochastic simulation / Ca imaging / Exocytosis / Systems biology
Research Abstract

Exocytosis is an important biological phenomenon, especially for neural and endocrine cells. Although several experimental techniques from various fields of sciences including biophysics, cell biology, biochemistry and also molecular biology have been developed for analyzing this phenomenon, we have not reached the quantitative understandings for exocytosis. We, therefore, developed a systems biological approach for exocytosis with the combinatorial method of biological imaging and stochastic simulation. For the quantitative simulation, simultaneous imaging for intracellular Ca increase as a trigger of exocytosis, and also for quantitative counting of events of exocytosis is required. An imaging method by a dual view technique enables us to visualize the intracellular Ca response by a fluorescent dye, Fura-red and exocytosis by GFP-fused neuropeptide Y. Stimulation by ATP induced a quick and transient response of Ca and following exocytosis with a several seconds delay. This indicates that we can trace the exocytosis from the first trigger response of intracellular Ca and the final event of neurotransmitter release in single cells. Next, we develop a new simulation technique based on the stochastic Gillespie method. Pre-fusion process of exocytosis was divided to 5 stages, and the transition probabilities were estimated to fit the experimental data We succeed to reveal the exocytosis induced by a transient intracellular Ca increase. This stochastic model includes the enzymatic activity for modulating exocytosis. Finally, we also succeed to simulate the sequential exocytosis reported in chromaffin cells with double cascade of the single stochastic model. We conclude that simultaneous imaging and quantitative stochastic simulation provide us a new analytical technique for systems biology of exocytosis.

  • Research Products

    (9 results)

All 2008 2007

All Journal Article (4 results) (of which Peer Reviewed: 2 results) Presentation (5 results)

  • [Journal Article] Dendritic design implements algorithm for synaptic extraction of sensory information2008

    • Author(s)
      H. Ogawa, G. I. Cummins, G. A. Jacobs and K. Oka
    • Journal Title

      The Journal of Neuroscience 28(18)

      Pages: 4592-4563

    • Description
      「研究成果報告書概要(和文)」より
    • Peer Reviewed
  • [Journal Article] Ca2+ influx through P2X receptors induces actin cytoskeleton reorganization by the formation of cofilin rods in neurites2008

    • Author(s)
      K. Homma, Y. Niino, K. Hotta and K. Oka
    • Journal Title

      Molecular and Cellular Neuroscience 37

      Pages: 261-270

    • Description
      「研究成果報告書概要(和文)」より
    • Peer Reviewed
  • [Journal Article] Dendritic design implements algorithm for synaptic extraction of sensory information2008

    • Author(s)
      H., Ogawa, G. I., Cummins, G. A., Jacobs, K., Oka
    • Journal Title

      The Journal of Neuroscience 28(18)

      Pages: 4593-4602

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Ca2+ influx through P2X receptors induces actin cytoskeleton reorganization by the formation of cofilin rods in neurites2008

    • Author(s)
      K., Homma, Y., Niino, K., Hotta, K., Oka
    • Journal Title

      Molecular and Cellular Neuroscience 37

      Pages: 261-270

    • Description
      「研究成果報告書概要(欧文)」より
  • [Presentation] Dual FRET imaging to visualize with two FRET sensors in signal cells2008

    • Author(s)
      Y., Niino, K., Hotta, K., Oka
    • Organizer
      45th Annual Meeting of the Biophysical Society of Japan
    • Place of Presentation
      Yokohama (Japan)
    • Year and Date
      20081222-23
    • Description
      「研究成果報告書概要(欧文)」より
  • [Presentation] Dual FRET imaging to visualize with two FRET probes simultaneously in signal cells2007

    • Author(s)
      Y. Niino, K. Hotta and K. Oka
    • Organizer
      第30回分子生物学会・第80回日本生化学学会合同大会
    • Place of Presentation
      パシフィコ横浜
    • Year and Date
      2007-12-14
    • Description
      「研究成果報告書概要(和文)」より
  • [Presentation] Dual FRET imaging to visualize with two FRET probes simultaneously in signal cells2007

    • Author(s)
      Y., Niino, K., Hotta, K., Oka
    • Organizer
      Biochemistry and Molecular Biology 2007
    • Place of Presentation
      Yokohama (Japan)
    • Year and Date
      2007-12-14
    • Description
      「研究成果報告書概要(欧文)」より
  • [Presentation] Dual FRET microscopy for imaging two intracellular signals simultaneously2007

    • Author(s)
      Y. Niino, K. Hotta and K. Oka
    • Organizer
      Society for Neuroscience 2007
    • Place of Presentation
      San Diego,USA
    • Year and Date
      2007-11-04
    • Description
      「研究成果報告書概要(和文)」より
  • [Presentation] Dual FRET microscopy for imaging two intracellular signals simultaneously2007

    • Author(s)
      Y., Niino, K., Hotta, K., Oka
    • Organizer
      Society for Neuroscience 2007
    • Place of Presentation
      San Diego(USA)
    • Year and Date
      2007-11-04
    • 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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