Regulation and effects of intracellular Cl concentrations

细胞内 Cl 浓度的调节和影响

• 批准号: 09470027
• 负责人: INAGAKI Chiyoko
• 金额: $ 6.91万
• 依托单位: Kansai Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09470027/
• 关键词: chloride ion; c-fos expression; brain; ammonia; anion exhanger; protein kinase C; hypoxia; chloride pump; クロライドイオン; c-fos; アミロイドβ蛋白; ナトリウム利尿ペプチド; エタクリン酸; 神経成長因子

(1) Intraneuronal ClィイD1=ィエD1 concentrations ([ClィイD1=ィエD1]i) and immediately early gene (c-fos) expressionIntracerebroventricular administration of a ClィイD1-ィエD1 pump inhibitor, ethacrynic acid, caused convulsions in mice. Expression of c-fos was stimulated biphasically with the peaks at 60min and 10-14 days after the convulsion. Expression of nerve growth factor, damage of GABAergic neurons and development of seizure susceptibility were also observed. A transient increase in neuronal [ClィイD1-ィエD1]i thus appeared to cause biphasic increases in c-fos expression and related modulation of neuronal functions.(2) Mechanisms of ammonia-induced increases in neuronal [ClィイD1=ィエD1]iA neurotoxic factor in hepatic encephalopathy, ammonia (2 mM, 48 hr), increased [ClィイD1-ィエD1]i in cultured rat hippocampal neurons. The increase was found to be due to enhanced expression of anion exchanger (AE3) mediated by protein kinase C activation.(3) ClィイD1=ィエD1 transport in hypoxia/reoxygenaion-induced changes in pHi and {CaィイD12+ィエD1]i in myocytesUnder hypoxia/reoxygenaion conditions, pHi decreased and [CaィイD12+ィエD1]i increased during hypoxia and reoxygenation, respectively. Inhibitors of anion exchanger (SITS and DIDS) and removal of medium ClィイD1-ィエD1 or HCOィイD23ィエD2 attenuated such changes in pHi and [CaィイD12+ィエD1]i. Inhibitors of protein kinase C slightly reduced the changes in pHi. Involvement of anion exchanger in hypoxia-induced pHi changes was first demonstrated with the resulting inhibitory effects on reoxygenation-induced increases in [CaィイD12+ィエD1]i.(4) Subunits and cDNA cloning of neuronal ClィイD1=ィエD1 pumpA new ClィイD1-ィエD1 transporter, ClィイD1-ィエD1 pump, was isolated as 520 kDa protein complex from the rat brain. SDS-PAGE analyses yielded 4 subunits (51, 55, 60 and 62kDa), and 51kDa protein appeared to be a catalytic subunit. cDNA for 55 kDa protein was cloned from the rat brain cDNA library, and its primary structure was assumed to be a new peptide with 475 amino acids.

（1）神经内神经元CLII D1 = IE D1浓度（[CLII D1 = IE D1] I）和立即的早期基因（C-FOS）表达CLII D1泵抑制剂乙酸乙酸的脑室内给药，导致小鼠中的抽搐。在抽搐后的60分钟和10-14天的峰值上刺激C-FOS的表达。还观察到神经生长因子的表达，GABA能神经元的损伤以及癫痫发作的发育。 A transient increase in neuronal [ClI D1-Ie D1]i thus appeared to cause biphasic increases in c-fos expression and related modulation of neuronal functions.(2) Mechanisms of ammonia-induced increases in neuronal [ClI D1=Ie D1]iA neurotoxic factor in hepatitic encephalopathy, ammonia (2 mM, 48 hr), increased [CLI D1-IE D1] I在培养的大鼠海马神经元中。发现增加是由于蛋白激酶C激活介导的阴离阴离子交换（AE3）的增强所致。（3）CLI D1 = IED1在低氧/reoxygenaion中的pHi和{CAI D12+IED1的变化中，在低氧/roxygenaion中均增加了phi和[CAI D12+IED1] I i EDD12+IC的IED12+CAII+CAII+CAII+CAII+[CAI ICRIP]+[CAI ICRIP]分别重氧。阴离子交换的抑制剂（位于DIDS）以及中位CLI D1-E D1或HCOI D23E D2的中位数抑制剂减弱了PHI和[CAI D12+E D1]中的此类变化。蛋白激酶C的抑制剂略微降低了PHI的变化。首先证明了阴离子交换器参与缺氧引起的PHI变化的参与，从而抑制了对重氧的抑制作用，对[CAI D12+E D1]I。（4）神经元CLI CLI D1 = E D1 pumpa New Cli d1 d1 pumper cli a kd kd a As cli autment，cli d1 = e d1 pumper的抑制作用，cai d12+e d1] i。 脑。 SDS-PAGE分析产生了4个亚基（51、55、60和62KDA），而51KDA蛋白似乎是催化亚基。从大鼠脑cDNA文库中克隆了55 kDa蛋白的cDNA，其主要结构被认为是具有475个氨基酸的新肽。

项目成果

Zeng, X.-T., Higashida, T., Hara, M., Hattori, N., Kitagawa, K., Omori, K. and Inagaki, C.: "Antiserum against ClィイD1-ィエD1 pump complex recognized 51 kDa protein, a posible catalytic unit in the rat brain."Neurosci. Lett.. 258. 85-88 (1998)

Zeng, X.-T.、Higashida, T.、Hara, M.、Hattori, N.、Kitakawa, K.、Omori, K. 和 Inagaki, C.：“针对 CliiD1-ieD1 泵复合物识别的 51 kDa 蛋白的抗血清，大鼠大脑中可能的催化单元。“Neurosci. Lett.. 258. 85-88 (1998)

稲垣千代子: "(Cl^-チャネルとCl^-輸送体) 「創薬」分担"共立出版(長尾拓他編). 5 (2000)

Chiyoko Inagaki：“（Cl^-通道和 Cl^-转运蛋白）‘药物发现’共享”Kyoritsu Shuppan（由 Taku Nagao 等人编辑）5 (2000)。

Yagyu, K. 他: "Lithium decreases Cl-ATPase activity and increases intracellular Cl concentration in cultured hippocampal neurons."Brain Res.. 821. 530-534 (1999)

Yagyu, K. 等人：“锂可降低培养的海马神经元中 Cl-ATP 酶活性并增加细胞内 Cl 浓度。”Brain Res.. 821. 530-534 (1999)

Minamino M, et al: "Effects of protein kinase and phosphatase inhibitons ・・" Brain Research. (in press). (1998)

Minamino M 等人：“蛋白激酶和磷酸酶抑制剂的影响··”大脑研究（出版中）。

Kawasaki E. et al.: "Single cell RT-PCR demonstrates expression of"Brain Research. 838. 166-170 (1999)

Kawasaki E. 等人：“单细胞 RT-PCR 证明了”Brain Research 的表达。