Affiliation: | a Graduate School of Human and Natural Environment Sciences, The University of Tokushima, Tokushima 770, Japan b Graduate School of Information Sciences, Tohoku University, Sendai 980-77, Japan c National Institute for Minamata Disease, Minamata 867, Japan |
Abstract: | 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. |