Tri- n-butyltin increases intracellular Zn 2+ concentration by decreasing cellular thiol content in rat thymocytes

Abstract

Effect of tri- n-butyltin (TBT), an environmental pollutant, on intracellular Zn 2+ concentration was tested in rat thymocytes to reveal one of cytotoxic profiles of TBT at nanomolar concentrations using a flow cytometer and appropriate fluorescent probes. TBT at concentrations of 30 nM or more (up to 300 nM) significantly increased the intensity of FluoZin-3 fluorescence, an indicator for intracellular Zn 2+ concentration, under external Ca 2+- and Zn 2+-free condition. Chelating intracellular Zn 2+ completely attenuated the TBT-induced augmentation of FluoZin-3 fluorescence. Result suggests that nanomolar TBT releases Zn 2+ from intracellular store site. Oxidative stress induced by hydrogen peroxide also increased the FluoZin-3 fluorescence intensity. The effects of TBT and hydrogen peroxide on the fluorescence were additive. TBT-induced changes in the fluorescence of FluoZin-3 and 5-chloromethylfluorescein, an indicator for cellular thiol content, were correlated with a coefficient of −0.962. Result suggests that the intracellular Zn 2+ release by TBT is associated with TBT-induced reduction of cellular thiol content. However, chelating intracellular Zn 2+ potentiated the cytotoxicity of TBT. Therefore, the TBT-induced increase in intracellular Zn 2+ concentration may be a type of stress responses to protect the cells.