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2001 Fiscal Year Final Research Report Summary

Development of Evanescence Microscope and Test of the Quantal Hypothesis for Exocytosis

Research Project

Project/Area Number 10557003
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section展開研究
Research Field General physiology
Research InstitutionHamamatsu University School of Medicine

Principal Investigator

TERAKAWA Susumu  Hamamatsu University School of Medicine, Photon Medical Research Center,, 光量子医学研究センター, 教授 (50014246)

Co-Investigator(Kenkyū-buntansha) ABE Katsuyuki  Olympus Optical Co., Research and Development Center, Researcher, 光学技術部・研究員
SAKURAI Takashi  Hamamatsu University School of Medicine, Photon Medical Research Center, Research Associate, 光量子医学研究センター, 助手 (50283362)
Project Period (FY) 1998 – 2000
Keywordswater secretion / exocytosis / evanescent microscope / Cl channel / laser trapping / insulin release / DIDS / ultra high NA objective lens
Research Abstract

Aiming at development of a user-friendly high-performance evanescence microscope based on an ultra-high NA lens, we compared three methods of introducing the illumination light to the microscope. A fiber-illumination method was advantageous in safety, field size, and easy handling, over a direct-illumination method, but was costly and slightly unstable. Using an evanescence microscope, we examined the exocytosis in chromaffin cells and beta cells. In both types of the cell, a fluorescent probe was released from vesicles with a flash response. The flash response was variable in amplitude depending on vesicles. It reflected water secretion from the vesicles. By laser-trap forcemetry, we demonstrated a small force exerted on water by the cell upon stimulation. The contents of vesicles are released not by a simply diffusion but by a water ejection from the vesicles. The strength of the ejecting force varied with vesicles as the density of Cl channels on the vesicle membrane examined by an immuno-staining technique varied. Cl channel blockers suppressed the flash response without suppressing the exocytosis. In beta cells, vesicle moved along the membrane immediately after its exocytotic response, suggesting a continuous release of vesicle contents. These findings are incompatible with the so-called quantal hypothesis. In endocrine cells, modulation for the secretory function is evolved in a highly complex manner so that fine adaptations to inner and outer environments are made possible.

  • Research Products

    (12 results)

All Other

All Publications (12 results)

  • [Publications] Ishihara Y: "Exocytosis and movement of zymogen granules observed by VEC-DIC microscopy in the pancreatic tissue en bloc."Am J Physiol (Cell Physiol). 279(4). C1177-C1188 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ishihara Y: "Exocytosis in the dissociated pancreatic acinar cells of the guinea pig directly visualized by VEC-DIC microscopy"Biochem Biophys Res Commun. 277. 134-137 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Tsuboi T: "Simultaneous evanescent wave imaging of insulin vesicle membrane and cargo during a single exocytonic event"Current Biology. 10. 1307-1310 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kawano Y: "High numerical aperture objective lenses and optical system improved objective type total internal reflection fluorescence microscopy"Proc SPIE. 4098. 142-151 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Naito K: "Calicheamicin-conjugated humanized anti-CD33 monoclonal antibody (gemtuzumab zogamicin, CMA-676) shows cytocidal effect on CD33-positive leukemia cell"Leukemia. 14. 1436-1443 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Tsuboi T: "Water Secretion Associated with Exocytosis in Endocrine Celsl Revealed by Micro Forcemetry and Evanescent Wave Microscopy"Biophys J. (in press). (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ishihara Y, Sakurai T, Kimura T, Terakawa S: "Exocytosis and movement of zymogen granules observed by VEC-DIC microscopy in the pancreatic tissue en bloc"Am J Physiol (Cell Physiol). 279(4). C1177-C1188 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ishihara Y, Sakurai T, Habara Y, Busik VJ, Kanno T, Terakawa S: "Exocytosis in the dissociated pancreatic acinar cells of the guinea pig directly visualized by VEC-DIC microscopy"Biochem Biophys Res Commun. 277. 134-137 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Tsuboi T, Zhao C, Terakawa S, Rutter GA: "Simultaneous evanescent wave imaging of insulin vesicle membrane and cargo during a single exocytotic event"Current Biology. 10. 1307-1310 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kawano Y, Abe C, Kaneda T, Aono Y, Abe K, Tamura K, Terakawa S: "High numerical aperture objective lenses and optical system improved objective type total internal reflection fluorescence microscopy"Proc. SPIE. 4098. 142-151 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Naito K, Takeshita A, Shigeno K, Nakamura S, Fujisawa S, Shinjo K, Yoshida H, Ohnishi K, Mori M, Terakawa S, Ohno R: "Calicheamicin-conjugated humanized anti-CD33 monoclonal antibody (gemtuzumab zogamicin, CMA-676) shows cytocidal effect on CD33-positive leukemia cell lines, but is inactive on P-glycoprotein-expressing sublines"Leukemia. 14. 1436-43 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Tsuboi T, Kikuta T, Sakurai T, Terakawa S: "Water Secretion Associated with Exocytosis in Endocrine Cells Revealed by Micro Forcemetry and Evanescent Wave Microscopy"Biophys J. in press.

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2003-09-17  

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