DISTINCTION OF THE MECHANISMS FOR EXOCYTOSIS BY SIMULTANEOUS MEASUREMENTS WITH EVANESCENCE METHOD AND CONFOCAL METHOD

消逝法和共焦法同时测量区分胞吐机制

• 批准号: 17390055
• 负责人: TERAKAWA Susumu
• 金额: $ 9.59万
• 依托单位: HAMAMATSU UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-17390055/
• 关键词: 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

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.

通过设计一种带有电扫描仪的光学器件，该器件能够为细胞更深的部分提供荧光照明，从而观察到 3 种不同类型细胞的分泌囊泡，并特别注意囊泡在胞吐作用之前的运动。消逝照明的穿透深度是根据由于盖玻片与标准盖玻片的重叠而引起的亮点的移动来测量的。在肾上腺素分泌细胞中，细胞外 pH 值的变化不会影响胞吐作用的闪光模式，表明囊泡内 pH 值的升高不是闪光反应的原因。在表达 GFP-胰岛素的 INS-1 细胞中，一些囊泡在胞吐闪光反应之前快速移位至细胞膜。我们发现随着深度的变化，这种易位发生在细胞膜上。在这些细胞中，很少看到胞吐作用后立即发生易位。在 MIN-6 细胞中，许多囊泡平行于细胞膜滑动长距离。这些囊泡很少经历胞吐闪光反应。根据这些观察，我们得出结论，闪光响应表明货物材料从囊泡中扩散。进一步开发能够在短时间内测量物体速度的显微光学系统是必要的，我们目前正在沿着这条线努力。

项目成果

光音響効果を用いたカテーテルシステム

利用光声效应的导管系统

• 发表时间: 2006

DLP式エバネッセンス顕微鏡

DLP型倏逝显微镜

• 发表时间: 2006

光スキャナ装置(共焦点光スキャナ検出装置,光スキャナおよびそれに用いられるニボウディスク)

光学扫描仪装置（共焦光学扫描仪检测装置、光学扫描仪以及其中使用的Nibo盘）

• 发表时间: 2007

観察装置

观察装置

• 发表时间: 2017

DLP式スリット光走査顕微鏡

DLP型狭缝光扫描显微镜

• 发表时间: 2006