2001 Fiscal Year Final Research Report Summary
Development of Evanescence Microscope and Test of the Quantal Hypothesis for Exocytosis
| 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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| Keywords | water secretion / exocytosis / evanescent microscope / Cl channel / laser trapping / insulin release / DIDS / ultra high NA objective lens |
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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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Research Products
(12 results)
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[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
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