| Project/Area Number |
17390055
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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 | 一般 |
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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, PROFESSOR (50014246)
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| Co-Investigator(Kenkyū-buntansha) |
SAKURAI Takashi HAMAMATSU UNIVERSITY SCHOOL OF MEDICINE, PHOTON MEDICAL RESEARCH CENTER, RESEARCH ASSOCIATE (50283362)
WAKAZONO Yoshihiko HAMAMATSU UNIVERSITY SCHOOL OF MEDICINE, PHOTON MEDICAL RESEARCH CENTER, INVESTIGATOR (90377755)
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| Project Period (FY) |
2005 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥14,980,000 (Direct Cost: ¥14,200,000、Indirect Cost: ¥780,000)
Fiscal Year 2007: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000)
Fiscal Year 2006: ¥6,800,000 (Direct Cost: ¥6,800,000)
Fiscal Year 2005: ¥4,800,000 (Direct Cost: ¥4,800,000)
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| Keywords | MIN-6 CELL / SECRETORY GRANULE / INSULIN RELEASE / CRITICAL ANGLE / ULTRA HIGH NA LENS / SLIDING MOVEMENT / INTRACELLULAR PH / EXOCYOTOSIS / エバネッセンス顕微鏡 / 斜光照明顕微鏡 / 干渉顕微鏡 / 開口放出 / ホルモン分泌 / 破骨細胞 / 近接場光顕微鏡 / エバネッセント光顕微鏡 / 全反射照明蛍光顕微鏡 / 共焦点蛍光顕微鏡 / エキソサイトーシス / 膜融合 / 細胞骨格 / 水分泌 / 開口数 / 全反射照明顕微鏡 / インスリン / 共焦点顕微鏡 / DMD |
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| Research Abstract |
By devising an optics with the galvano-scanner that provides a capability of fluorescence illumination to a deeper part of cells than the evanescence microscopy, we observed secretory vesicles of 3 different types of cells with a special attention to the movement of the vesicles before exocytosis. The penetration depths of the evanescence illuminations were measured from the shift of the bright spot due to overlaying of a coverslip to the standard coverslip. In adrenaline secreting cells, a change in extracellular pH was not affecting to the flash pattern of the exocytosis, indicating that the rise in pH inside vesicles is not cause for the flash response. In INS-1 cells, expressing GFP-insulin, some vesicles translocated rapidly to the cell membrane before exocytotic flash response. We found such a translocation occurred toward the cell membrane as depth change. In these cells, it was rare to see translocations immediately followed by exocytosis. In MIN-6 cells, many vesicles glided for long distances in parallel to the cell membrane. These vesicles rarely underwent the exocytotic flash response. From these observations, we concluded that the flash response indicates the diffusion of cargo materials from vesicles. A further development of microscopic optics that can measure the velocity of object in short time would be necessary, and we are currently working along this line.
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