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Cl^-homeostasis failure and abnormal cell migration by endocrine disruptor cause damage to nervous system

内分泌干扰物导致的Cl^-稳态失灵和异常细胞迁移导致神经系统损伤

基本信息

批准号：
15590207
负责人：
YAMADA Junko
金额：
$ 2.11万
依托单位：
Shizuoka 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

项目摘要

Measurement of [Cl^-]i and cell miglationUsing in vivo electropolation, we transfected eGFP prasmid embryonic day 14 (E14) mice. After electropolation, mice were exposed tributyltin chloride (TBTCl ; 1μl) via amniotic fluid. 3-4 days after exposure TBTCl, mice were observed their conditions.Results were as follows ;1.TBTCl 1M : Mice were dead with atrophy. Both side of mice, which were intact, also dead.2.500 mM-100 mM : Mice were dead with atrophy. Both side of mice, which were intact, survived.3.10 mM : Mice were dead but looks were normal.4.1 mM : Mice sere survived and cell miglation was normal.The effect of TBTCl exposure on Cl^-homeostasis.Using gramicidin perforate patch-clamp method, we measured [Cl^-]_i TBTCl exposed cell (30 nM, 3days) and intact cell (0.1 % DMSO, 3 days). Both cell group had normal [Cl^-]_i.The effect of TBTCl exposure on GABA receptor expressionmRNAs of TBTCl exposed cell and control cell were harvested. Using RT-PCR method, expression of GABAA receptor subunit (α1-5、β1-3、γ1-3、δ、ε、and β-actin)were analysed.The acute effect of TBTCl on culture cell.Using patch-clamp method and Ca2+ imaging, neurons were observed effect of TBTCl application. At low concentration of TBTCl application (30 nM), there were no effect on membrane potantial and synaptic current. It seems that toxicity at low concentration of TBTCl acts slowly.

[Cl~(-)]i的测定和细胞迁移利用体内电极法，将EGFP基因导入胚胎第14天(E14)小鼠。电极化后，小鼠经羊水暴露三丁基氯化锡(1μL)。染毒后3-4天，观察小鼠的情况，结果如下：1.四氯化碳1M：小鼠萎缩死亡。小鼠双侧完好，也死亡。2.500 mm-100 mm：小鼠萎缩死亡。小鼠双侧完好存活。3.10 mm：小鼠死亡，但外观正常。4.1 mm：小鼠存活，细胞迁移正常。四叔丁基氯暴露对氯离子稳态的影响。采用革兰西丁穿孔膜片钳方法，测定[Cl2+]_i TBTCl暴露细胞(30 nM，3天)和完整细胞(0.1%DMSO，3天)。两组细胞的[Cl~-]_i均正常。观察了TBTCl暴露对细胞内GABA受体表达的影响。采用逆转录聚合酶链式反应的方法分析α1-5、、β1-3、、γ1-3、、δ、ε、和β-肌动蛋白受体亚单位的表达。低浓度的三氯甲烷(30 NM)对膜电位和突触电流无明显影响。低浓度的四丁基氯的毒性作用似乎较慢。

项目成果

期刊论文数量（42）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Transgenic Expression Enhanced Green Fluorescent Protein Enables Direct Visualization for Physiological Studies of Vasoressin Neurons and Isolated Nerve Terminals of the Rat.

转基因表达增强的绿色荧光蛋白使得血管加压素神经元和大鼠离体神经末梢的生理学研究能够直接可视化。

DOI：
发表时间：
2004
期刊：
Endocrinology. 24
影响因子：
0
作者：
Ueta;Y.;Fujihara;H.;Yamada et al.
通讯作者：
Yamada et al.

Cl- uptake promoting depolarizing GABA actions in immature rat neocortical neurones is mediated by NKCC1

DOI：
10.1113/jphysiol.2004.062471
发表时间：
2004-06-15
期刊：
JOURNAL OF PHYSIOLOGY-LONDON
影响因子：
5.5
作者：
Yamada, J;Okabe, A;Fukuda, A
通讯作者：
Fukuda, A

Neuroprotective effects of melatonin against anoxia/aglycemia stress, as assessed by synaptic potentials and superoxideproduction in rat hippoocampal slices

通过大鼠海马切片的突触电位和超氧化物产生评估褪黑素对缺氧/血糖应激的神经保护作用

DOI：
发表时间：
2004
期刊：
J.Pineal Res. 37
影响因子：
0
作者：
Uchida K.;Samejima M.;Okabe A.;Fukuda A.
通讯作者：
Fukuda A.

Experience-dependent changes in intracellular Cl- regulation in developing auditory neurons

DOI：
10.1016/j.neures.2003.10.011
发表时间：
2004-02-01
期刊：
NEUROSCIENCE RESEARCH
影响因子：
2.9
作者：
Shibata, S;Kakazu, Y;Nabekura, J
通讯作者：
Nabekura, J

Malfunction of respiratory related neuronal activity in Na^+, K^+-ATPase α2 subunit-deficient mice is due to abnormal Cl^- homeostasis in brainstem neurons.

Na^+、K^+-ATPase α2 亚基缺陷小鼠呼吸相关神经元活动功能障碍是由于脑干神经元中 Cl^- 稳态异常所致。

DOI：
发表时间：
2004
期刊：
J.Neurosci. 24
影响因子：
0
作者：
Ban;T.et al.;Toyoda H. et al.;Ueta Y.et al.;Inoue K. et al.;Wang C.et al.;Inoue K. et al.;Yamada J. et al.;Uchida K. et al.;Ikeda K. et al.
通讯作者：
Ikeda K. et al.