| Project/Area Number |
10557003
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
General physiology
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| Research Institution | Hamamatsu University School of Medicine |
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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)
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| 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)
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| Project Period (FY) |
1998 – 2000
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥11,800,000 (Direct Cost: ¥11,800,000)
Fiscal Year 2000: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1999: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1998: ¥7,100,000 (Direct Cost: ¥7,100,000)
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| Keywords | water secretion / exocytosis / evanescent microscope / Cl channel / laser trapping / insulin release / DIDS / ultra high NA objective lens / 全反射照明蛍光顕微鏡 / 一分子蛍光 / ホルモン / 膜融合 / 嚢胞性線維症 / エキソサイトーシス / 量子仮説 / 全反射照明 / 神経伝達物質 / 高開口数レンズ / Clチャンネル / 全反射顕微鏡 / エバネッセンス / クロマフィン細胞 / 開口数 |
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| 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.
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