Concerted mechanisms of transporters for exocrine secretion

该项目旨在研究外分泌腺泡细胞中转运蛋白(通道、共转运、反向运输和泵)产生宏观分泌(协调机制)的协同机制。该项目的重点是唾液腺中转运蛋白的运输能力、能量供应及其控制。合作实验室使用不同的专门技术，测量了分离的细胞和血管灌流的整个腺体中跨细胞膜和细胞内信息系统的水和电解质的移动。NIPS小组开发了一种分离和灌流腮腺的技术。他们同时测量液体分泌和氧气消耗，并评估钠依赖的转运体对有组织分泌的贡献。曼彻斯特/冈崎研究小组使用脉冲梯度核磁共振和荧光法测量了整个腺体和分离的细胞的细胞体积变化。他们发现细胞内氯与细胞体积呈正相关。松岛队Fo…兔腮腺内cGMP积聚与一氧化氮合酶活性有关。他们还发现VAMP-2是囊泡膜中的一种胞吐相关蛋白。哥本哈根团队测量了肌醇化合物在分泌过程中的时间进程，发现嘌呤能受体的激活抑制了物质P对IP3的动员。NIPS/哈佛团队观察到ATP抑制ACh的液体分泌，这在实验上得到了支持。Harverd研究小组发现，CCH和P物质作为早期事件引起PKC Delta的磷酸化，然后MAP激酶被激活。悉尼研究小组研究了导管分泌钾和钠重吸收的机制。他们得出结论，Na/K/2Cl共转运对小鼠导管K的分泌没有贡献，并发现G蛋白参与了导管对氯化钠的重吸收。NIH课题组从分子水平研究了Na/K/2Cl共转运蛋白的激活机制，发现该共转运蛋白可能对唾液中的铵进行生理性排泄。Kitasato/Cagliari团队观察了细胞间路径和胞吐作用的3D结构，并发现CCH和异丙肾上腺素刺激下通过微丝的胞吐-内吞耦合作用存在功能差异。合作使每个实验室可以使用相同的材料制备技术和截然不同的测量技术，因此产生了实质性的协同结果。较少

The project aims to study a co-operative mechanism of the transporter proteins (channel, cotransport, antiport & pumps) to produce a macroscopic secretion (concerted mechanism) in the exocrine acinar cells. The project focuses on transport ability, energy supply and its control of transporter proteins in the salivary gland. Collaborative laboratories, using different specialized techniques, measured water & electrolyte movements across cell membranes and intracellular information system in the isolated cells and the vascularly perfused whole gland. NIPS group developed a technique to isolate and perfuse the parotid gland. They measured fluid secretion and oxygen consumption simultaneously, and assessed contribution of Na-dependent transporters to organized secretion. Manchester/Okazaki team measured cell volume change in the whole gland and isolated cells using pulse gradient NMR and fluorometry. They found positive relationship between intracellular Cl and cell volume. Matsudo team fo … More und an activity of NOS involving cGMP accumulation in the rabbit parotid gland. They also identified VAMP-2 as a exocytosis-related protein in the vesicle membrane. Copenhagen team measured inositol compounds along the time course of secretion, and found that activation of purinergic receptor inhibit mobilization of IP3 by substance P.NIPS/Harvard team supported experimentally by observation that ATP inhibits fluid secretion by ACh. Harverd team found that CCh and substance P caused phosphorylation of PKC delta as early event, then MAP kinase was activated. Sydney group examined mechanism of ductal K secretion and Na reabsorption. They concluded no contribution of Na/K/2Cl cotransport for ductal K secretion in the mouse, and found involvement of G-protein on NaCl reabsorption in the duct. NIH group examined activation mechanism of Na/K/2Cl cotransporter at molecular level, and found a possible role of cotransporter to excrete ammonium physiologically from saliva. Kitasato/Cagliari team observed 3D structure of intercellular path and exocytosis, and found functional difference in the exocytosis-endocytosis coupling via microfilament between CCh and isoproterenol stimulation. The collaboration allowed to use the same preparation technique of the materials and quite different measurement techniques used by each laboratories, so substantial synergetic results was produced. Less