The effects of tri-n-butyltin on cell viability and intracellular Ca2+ concentration ([CA2+]i) were examined by a flow cytometer in rat cerebellar neurones to reveal the contribution of tri-n-butyltin-induced increased in the [Ca2+]i to its cytotoxicity. Tri-n-Butyltin decreased the cell viability in association with increased [Ca2+]i at concentrations of 0.3 microM or more. Decrease or increase of the extracellular Ca2+ concentration ([Ca2+]o) respectively decreased or increased the cell viability. However, cell viability in the presence of ionomycin which increased [Ca2+]i more significantly than tri-n-butyltin was higher than that in the presence of tri-n-butyltin. Tri-n-butyltin also decreased cell viability under nominally [Ca2+]o-free conditions although the [Ca2+]i increase by tri-n-butyltin was still lower than the control [Ca2+]i under normal [Ca2+]o (2 mM). Therefore, it is unlikely that neuronal death induced by tri-n-butyltin is entirely dependent on the tri-n-butyltin-induced increase in [Ca2+]i.