TY - JOUR
T1 - Fluorescent estimation on cytotoxicity of methylmercury in dissociated rat cerebellar neurons
T2 - Its comparison with ionomycin
AU - Okazaki, Eisuke
AU - Oyama, Yasuo
AU - Chikahisa, Lumi
AU - Nagano, Takayuki
AU - Katayama, Norihiro
AU - Sakamoto, Mineshi
N1 - Funding Information:
This study was supported by a research grant to Y. Oyama from the Japan Public Health Association, Tokyo, Japan.
PY - 1997/9
Y1 - 1997/9
N2 - 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.
AB - 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.
KW - Cell membrane
KW - Cerebellar neurons
KW - Flow cytometer
KW - Fluorescent dyes
KW - Intercellular Ca
KW - Methylmercury
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U2 - 10.1016/S1382-6689(97)00017-3
DO - 10.1016/S1382-6689(97)00017-3
M3 - Article
C2 - 21781783
AN - SCOPUS:0030880491
VL - 3
SP - 237
EP - 244
JO - Environmental Toxicology and Pharmacology
JF - Environmental Toxicology and Pharmacology
SN - 1382-6689
IS - 4
ER -