Concerted mechanisms of transporters for exocrine secretion

本项目旨在研究外分泌腺泡细胞中转运蛋白（通道、共转运、反转运和泵）协同分泌的机制（协同机制）。该项目重点研究唾液腺转运能力、能量供应及其对转运蛋白的控制。合作实验室使用不同的专业技术，测量了水和电解质在分离细胞和血管灌注的整个腺体中跨细胞膜和细胞内信息系统的运动。NIPS组开发了腮腺分离灌注技术。他们同时测量了液体分泌和氧气消耗，并评估了na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与细胞体积呈正相关。Matsudo团队在兔腮腺中发现了与cGMP积累有关的NOS活性。他们还发现VAMP-2是囊泡膜中的胞吐相关蛋白。哥本哈根团队沿着分泌的时间过程测量了肌醇化合物，发现嘌呤能受体的激活抑制了p物质对IP3的动员。nips /Harvard团队通过观察ATP抑制ACh分泌液体的实验支持了这一观点。哈佛大学的研究小组发现，CCh和P物质作为早期事件引起PKC三角洲磷酸化，然后MAP激酶被激活。Sydney组检测了导管K分泌和Na重吸收的机制。他们得出结论，Na/K/2Cl共转运对小鼠导管中K的分泌没有贡献，并发现g蛋白参与了导管中NaCl的重吸收。NIH组在分子水平上考察了Na/K/2Cl共转运体的激活机制，发现了Na/K/2Cl共转运体可能在唾液中生理排泄铵的作用。Kitasato/Cagliari团队观察了胞间通路和胞吐的三维结构，并发现CCh和异丙肾上腺素刺激在胞吐-胞吞耦合的微丝功能上存在差异。这次合作允许每个实验室使用相同的材料制备技术和完全不同的测量技术，因此产生了大量的协同结果。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