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

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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.