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Analysis of endosome/post-endosome system with special reference to water channel aquaporin 2

特别参考水通道水通道蛋白 2 的内体/内体后系统分析

基本信息

批准号：
18390057
负责人：
TAKATA Kuniaki
金额：
$ 10.21万
依托单位：
Gunma University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2009
项目状态：
已结题

项目摘要

Vesicular trafficking that determines the number of cell surface molecules serves as one of important regulatory mechanisms found in the control of the reabsorption of water in the kidney, maintenance of blood glucose level, etc.Intracellular vesicles involved in the endocytosis and subsequent recycling process, such as endosomes and post-endosomal vesicles are key components in such vesicular trafficking and storage. We used water channel aquaporin 2 whose trafficking is regulated by hypophysial hormone vasopressin. Glucose transporter 4(GLUT4), whose trafficking is regulated by insulin in adipocytes and muscle cells, was coexpressed with aquaporin 2 in MDCK cells, and their localization was visualized by immunofluorescence and GFP-tagging methods. Aquaporin 2 and GLUT4 exhibited distinct localization pattern each other in both the resting and stimulated states, showing that aquaporin 2 and GLUT4 are stored in distinct post-endosomal compartments each other, and that their trafficking is differentially regulated. Translocation of aquaporin 2 has been considered to be regulated by its phosphorylation. We raised antibodies that specifically detect phosphorylated aquaporin 2. By western blotting and immunofluorescence microscopy, phosphorylation of aquaporin 2 was found not only on the cell surface but also intracellular vesicles, suggesting the ubiquitous presence of phosphorylated aquaporin 2.

囊泡运输决定了细胞表面分子的数量，是控制肾脏水重吸收、维持血糖水平等的重要调节机制之一。参与内吞和随后再循环过程的细胞内囊泡，如内体和内体后囊泡，是囊泡运输和储存的关键组分。我们使用水通道水通道蛋白2，其运输受垂体激素加压素调节。葡萄糖转运蛋白4（GLUT4），其运输在脂肪细胞和肌肉细胞中由胰岛素调节，在MDCK细胞中与水通道蛋白2共表达，并通过免疫荧光和GFP标记方法观察其定位。水通道蛋白2和GLUT 4在静息和刺激状态下均表现出不同的定位模式，表明水通道蛋白2和GLUT 4储存在彼此不同的内体后区室中，并且它们的运输受到差异调节。水通道蛋白2的转运被认为是由其磷酸化调节的。我们提出了特异性检测磷酸化水通道蛋白2的抗体。通过免疫印迹和免疫荧光显微镜，磷酸化的水通道蛋白2被发现不仅在细胞表面，但也细胞内囊泡，这表明磷酸化的水通道蛋白2的普遍存在。

项目成果

期刊论文数量（0）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Immunohistochemical localization of the aquaporins AQP1, AQP3, AQP4, and AQP5 in the mouse respiratory system.

DOI：
10.1267/ahc.09023
发表时间：
2009-12-29
期刊：
Acta histochemica et cytochemica
影响因子：
2.4
作者：
Matsuzaki T;Hata H;Ozawa H;Takata K
通讯作者：
Takata K

Localization and trafficking of aquaporin 2 in the kidney.

DOI：
10.1007/s00418-008-0457-0
发表时间：
2008-08
期刊：
HISTOCHEMISTRY AND CELL BIOLOGY
影响因子：
2.3
作者：
Takata, Kuniaki;Matsuzaki, Toshiyuki;Tajika, Yuki;Ablimit, Abduxukur;Hasegawa, Takahiro
通讯作者：
Hasegawa, Takahiro

免疫シナプスにおける分子分布動態の3次元ライブセルイメージング解析

免疫突触分子分布动态的 3D 活细胞成像分析