Concerted mechanisms of transporters for exocrine secretion

该项目旨在研究转运蛋白（通道、共转运、反转运和泵）的​​协作机制，以在外分泌腺泡细胞中产生宏观分泌（协同机制）。该项目的重点是唾液腺中转运蛋白的转运能力、能量供应及其控制。合作实验室使用不同的专业技术，测量了分离细胞和血管灌注的整个腺体中水和电解质跨细胞膜和细胞内信息系统的运动。 NIPS 小组开发了一种分离和灌注腮腺的技术。他们同时测量了液体分泌和耗氧量，并评估了钠依赖性转运蛋白对有组织分泌的贡献。曼彻斯特/冈崎团队使用脉冲梯度核磁共振和荧光测定法测量了整个腺体和分离细胞的细胞体积变化。他们发现细胞内 Cl 与细胞体积之间呈正相关。 Matsudo 团队发现 NOS 的活动涉及兔腮腺中 cGMP 的积累。他们还鉴定出 VAMP-2 是囊泡膜中的胞吐作用相关蛋白。哥本哈根团队测量了分泌时间过程中的肌醇化合物，发现嘌呤能受体的激活抑制了P物质对IP3的动员。NIPS/哈佛团队通过观察到ATP抑制ACh的液体分泌得到了实验支持。哈佛团队发现CCh和P物质作为早期事件引起PKC delta磷酸化，然后MAP激酶被激活。悉尼组检查了导管钾分泌和钠重吸收的机制。他们得出结论，Na/K/2Cl 共转运对小鼠导管 K 分泌没有贡献，并发现 G 蛋白参与导管中 NaCl 重吸收。 NIH小组在分子水平上研究了Na/K/2Cl协同转运蛋白的激活机制，发现协同转运蛋白可能具有从唾液中生理性排泄铵的作用。 Kitasato/Cagliari团队观察了细胞间路径和胞吐作用的3D结构，发现CCh和异丙肾上腺素刺激之间通过微丝的胞吐作用-内吞作用耦合的功能差异。此次合作允许使用相同的材​​料制备技术和每个实验室使用的截然不同的测量技术，因此产生了实质性的协同结果。较少的

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