Dynamic analysis by confocal microscopy of Salivary water secretion

唾液水分泌的共聚焦显微镜动态分析

• 批准号: 07671986
• 负责人: SEGAWA Akihisa
• 金额: $ 1.54万
• 依托单位: KITASATO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07671986/
• 关键词: Secretion; confocal microscopy; salivary glands; 水分泌; レーザー顕微鏡

Secretion mechanism of water by saliary glands was analyzed in living cells by confocal laser scanning microscopy. Tissue slices or dissociated acini were perfused with medium containing various fluorescent probes, and stimulated fluid secretion by adding carbachol. 1) membrane dynamics : The luminal membrane of acinar cells exhibited the long-term (more than 30 min) exocytosis-endocytosis coupling. Scanning and transmission electron microscopy revealed this system to be coated vesicle non-mediated. 2) Tight junction : Fluid-phase tracer dextrans-FITC and membrane marker FM1-43 revealed that dextrans up to MW 10K appeared in the lumen after the secretory stimulation whereas FM1-43 did not stain the luminal membrane, indicating that during fluid secretion tight junction opens its 'gate' but keeps its 'fence' function to remain unchanged. 3) IP3 receptors and Ca signaling Real-time Ca imaging by confocal microscopy exhibited Ca release from the apical cytoplasm of certain duct cells. Immunoelectron microscopy showed the IP3-reactive sites to be localized at the apical vesicles, which are considered newly defined IP3-receptor sites involved in Ca signaling.

用激光共聚焦显微镜观察唾液腺分泌水的机制。组织切片或解离腺泡灌注含有各种荧光探针的培养基，并通过加入卡巴胆碱刺激液体分泌。1)膜动力学：腺泡细胞腔膜呈现出长期（30 min以上）的胞吐-胞吞偶联。扫描和透射电子显微镜显示该系统被非介导的囊泡包被。2)紧密连接：液相示踪剂dextrans-FITC和膜标记物FM 1 -43显示，在分泌刺激后，高达MW 10 K的dextranss出现在管腔中，而FM 1 -43不染色管腔膜，表明在液体分泌期间，紧密连接打开其“门”，但保持其“栅栏”功能不变。3)IP 3受体和Ca信号传导通过共聚焦显微镜进行的实时Ca成像显示某些导管细胞的顶端细胞质中有Ca释放。免疫电子显微镜显示IP 3反应位点被定位在顶端囊泡，这被认为是新定义的IP 3受体位点参与钙信号。

项目成果

Akihisa Segawa: "Dynamic observation of all surface (in Japanese)" Saibou Kogaku (cell Technology). 15. 1159-1165 (1996)

Akihisa Sekawa：“所有表面的动态观察（日语）”Saibou Kogaku（细胞技术）。

A. Segawa & A. Rina: "Dynamics of Salivary Secretion stugied by Contocal Laser and Scanning Electron Hicroscopy" European Journal of Morphology. (印刷中). (1996)

A. Sekawa 和 A. Rina：“通过激光和扫描电子显微镜研究唾液分泌的动力学”《欧洲形态学杂志》（1996 年出版）。

瀬川彰久: "細胞表面の動的観察" 細胞工学. 15巻8号. 1159-1165 (1996)

濑川明久：“细胞表面的动态观察”《细胞工程》第 15 卷，第 8 期。1159-1165（1996 年）

Akihisa Sagawa: "Dynamicsof Salway Secretion studied by Confocal daser and Scanning Electrin Uicroscopy" European Journal of Mosphology. 34. 215-219 (1996)

佐川明久：“通过共焦激光和扫描电子显微镜研究萨尔韦分泌的动力学”欧洲形态学杂志。

Akihisa Segawa, Alessandro Rira: "Dynamics of Salivary Secretion studied by Confocal Laser and Scanning Electron Microscopy" European Journal of Morphology. 34. 215-219 (1996)

Akihisa Sekawa、Alessandro Rira：“通过共聚焦激光和扫描电子显微镜研究唾液分泌的动力学”欧洲形态学杂志。