The regulatory mechanism of amino acid release form swollen cells by aquaporins

水通道蛋白对肿胀细胞氨基酸释放的调控机制

• 批准号: 15590202
• 负责人: HAZAMA Akihiro
• 金额: $ 2.3万
• 依托单位: Fukushima Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15590202/
• 关键词: glutamamic acid; aquanorin; glia; Cl-channel; cell swelling; グリオーマ; C6; CL-チャンネル; グルコース; アミノ酸; 低浸透圧

We investigate the mechanism of amino acid release from swollen cells induced by the metabolic inhibition. Among the amino acids, we focused on the glutamate because of the importance in the brain anoxia. First we developed the method of detecting glutamate. We used the tetrazolium salts (WST-8, DOJIN) and glutamate dehydrogenase to form the formazan. With this combination, we could measure the concentration of glutamate using the plate reader. We applied the NaN3 to the glioma cell line (C6) and measured the glutamate in the extracellular solution. Extracellular gutamate increased from 3μM in the initial condition to 10μM after 30min. The rise of glutamate could not be suppressed by chloride channel blockers. We also tested HgC12 which could block aquaporin water pathway. By the application of HgC12, we could observed the rise of glutamate even without the metabolic inhibitor. This effect might be explained by the damage of cell membrane by HgC12. During this project, we also found the glutamate released form C6 cells by the application of extracellular glucose. This finding might contribute the investigation of mechanism of tissue damage caused by the high plasma glucose.

我们研究了代谢抑制引起的肿胀细胞释放氨基酸的机制，因为在大脑缺氧中的重要性，我们首先开发了检测谷氨酸的方法。 （WST-8，DOJIN）和谷氨酸脱氢酶形成甲臜，我们可以使用酶标仪测量谷氨酸的浓度。将 NaN3 加入到神经胶质瘤细胞系 (C6) 中，并测量细胞外溶液中的谷氨酸从初始条件下的 3μM 增加到 30 分钟后的 10μM。我们还测试了 HgC12。可以阻断水通道蛋白水途径 通过应用 HgC12，即使没有代谢抑制剂，我们也可以观察到谷氨酸的升高。这种效应可能是通过HgC12对细胞膜的损伤来解释的。在这个项目中，我们还发现了细胞外液从C6细胞中释放出的谷氨酸，这一发现可能有助于研究高血糖引起的组织损伤的机制。葡萄糖。

项目成果

Detecting ATP release by a biosensor method

• DOI: 10.1126/stke
• 发表时间: 2004-01-01
• 期刊: Sci. STKE
• 作者: Hayashi, S;Hazama, A;Okada, Y
• 通讯作者: Okada, Y