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Molecular and cellular morphological analysis of sugar transporter dynamics between cell membrane and organelles

细胞膜和细胞器之间糖转运蛋白动力学的分子和细胞形态学分析

基本信息

批准号：
15390053
负责人：
TAKATA Kuniaki
金额：
$ 9.86万
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：
2003
资助国家：
日本
起止时间：
2003 至 2005
项目状态：
已结题

项目摘要

Sugar transporter GLUT4 is recruited from the intracellular compartments and is translocated to the cell membrane upon insulin stimulation, which increases the sugar transport activity at the cell surface. GLUT4, therefore, plays a pivotal role in the regulation of blood glucose level. I first examined the localization of GLUT4 in 3T3-L1 cultured adipocytes by immunofluorescence microscopy. Insulin-stimulation mobilized only a small portion of intracellular GLUT4. The paucity of translocation of GLUT4 was a serious drawback in analyzing the mechanism of transporter trafficking by morphological methods. Aquaporin 2 water channel is also translocated from the intracellular pool to the cell membrane upon anti-diuretic hormone-stimulation in principal cells of the kidney collecting ducts, which increases the water permeability at the apical cell membrane. This system provides a unique opportunity to study the translocation mechanism between intracellular compartments and cell membrane. I examined the immunolocalization and trafficking of aquaporin 2 with special reference to intracellular organelles and microdomains of the embrane such as rafts. Aquaporin 2 was recovered in Triton X100-insoluble fractions, suggesting the contribution of membrane rafts. Stimulation with forskolin induces the translocation of aquaporin 2 to the cell membrane, where it was colocalized with caveolin 1. Wash-out of forskolin initiated the retrieval of aquaporin 2 to the EEA1-positive early endosomes by endocytosis. Such trafficking was affected by drugs that perturb membrane microdomains, suggesting an important role of membrane microdomains in the trafficking of aquaporin 2.

糖转运蛋白GLUT 4从细胞内区室募集，并在胰岛素刺激后易位至细胞膜，这增加了细胞表面的糖转运活性。因此，GLUT 4在血糖水平的调节中起着关键作用。我首先通过免疫荧光显微镜检查了GLUT 4在3 T3-L1培养的脂肪细胞中的定位。胰岛素刺激仅动员了一小部分细胞内GLUT 4。GLUT 4转运的缺乏是用形态学方法分析转运体转运机制的一个严重缺陷。在肾集合管的主细胞中，水通道蛋白2水通道也在抗利尿剂刺激后从细胞内池移位到细胞膜，这增加了顶端细胞膜处的水渗透性。该系统提供了一个独特的机会，研究细胞内室和细胞膜之间的易位机制。我研究了水通道蛋白2的免疫定位和运输，特别是细胞内细胞器和膜的微区，如筏。在Triton X100不溶性组分中回收水通道蛋白2，表明膜筏的贡献。用毛喉素刺激诱导水通道蛋白2易位到细胞膜，在那里它与小窝蛋白1共定位。冲洗毛喉素启动检索水通道蛋白2的EEA 1阳性早期内体的内吞作用。这种贩运受到药物的影响，扰乱膜微区，这表明水通道蛋白2的贩运膜微区的重要作用。

项目成果

期刊论文数量（44）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Sugar transporters and water channels in the Golgi apparatus and cell membrane. (in Japanese)

高尔基体和细胞膜中的糖转运蛋白和水通道。

DOI：
发表时间：
2005
期刊：
J Electron Microsc Technol Med Biol 19
影响因子：
0
作者：
Matsuzaki T;Tajika Y;Matsuzaki T;Morishita Y;Takata K;高田邦昭;Matsuzaki T;Tajika Y;Matsuzaki T;Takata K
通讯作者：
Takata K

Molecular mechanisms and drug development in aquaporin water chanel diseases : Water channel aquapori2 (AQP2) of kidney collecting duct cells.

水通道蛋白水通道疾病的分子机制和药物开发：肾集合管细胞的水通道 aquapori2 (AQP2)。

DOI：
发表时间：
2004
期刊：
J Pharmacol Sci 96
影响因子：
0
作者：
Matsuzaki T;Tajika Y;Matsuzaki T;Morishita Y;Takata K;高田邦昭;Matsuzaki T;Tajika Y;Matsuzaki T;Takata K;Tajika Y;Matsuzaki T;Morishita Y;Takata K;Takata K;Takata K
通讯作者：
Takata K

Aquaporin-2 is retrieved to the apical storage compartment via early endosomes and phosphatidylinositol 3-kinase-dependent pathway