Fluorescent estimation on cytotoxicity of methylmercury in dissociated rat cerebellar neurons: Its comparison with ionomycin

To study the cellular basis of the neurotoxicity of methylmercury, the effects of methylmercury on dissociated rat cerebellar neurons were examined using a flow cytometer, a confocal laser microscope and three fluorescent dyes, fluo-3 for monitoring the changes in intracellular Ca²⁺ concentration ([Ca²⁺](i)) and for detecting live neurons, ethidium for assessing the neurons that are dead or have compromised membranes, and 5-chloromethylfluorescein (CMF) for estimating the cellular content of nonprotein thiols. Methylmercury at concentrations of 1 μM or greater increased the [Ca²⁺](i) of almost all neurons. Prolonged exposure to methylmercury (3 and 10 μM) produced a further increase in [Ca²⁺](i), in association with compromising membranes in some neurons. Thereafter, methylmercury induced blebs on membranes of some neurons with increased [Ca²⁺](i). Methylmercury at concentrations of 0.3 μM or greater dose-dependently decreased the cellular content of nonprotein thiols. Results suggest that methylmercury may induce the loss of membrane integrity through destabilized Ca²⁺ homeostasis and oxidative stress in mammalian brain neurons.