KMID : 0043320180410111098
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Archives of Pharmacal Research 2018 Volume.41 No. 11 p.1098 ~ p.1107
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Sensitive neurotoxicity assessment of bisphenol A using double immunocytochemistry of DCX and MAP2
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Cho Jung-Hyun
Kim Ah-Hyun Lee Seul-Ah Lee Yu-Jeong Lee Won-Jong Chang Seung-Cheol Lee Jae-Won
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Abstract
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Bisphenol A (BPA) is an environmental toxin widely used in manufacturing industries. Studies conducted on the neurotoxicity of BPA demonstrated that at excessive, high concentrations (¡Ã?200 ¥ìM) adverse responses occurred which were not detectable using traditional toxicity tests at lower chemical quantities than 200 ¥ìM. Thus, a method capable of effectively detecting neurotoxicity at low concentrations (¡Â?100 ¥ìM) was devised. Bisphenol A-mediated neurotoxicity was examined in primary cultured neurons using various methods, including Western blot, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cytotoxicity and reactive oxygen species assays. These methods confirmed BPA-induced toxicity at 200 ¥ìM, but no marked effect was observed at concentrations below 200 ¥ìM. However, when immunocytochemistry (ICC) was performed using a co-immunofluorescence assay of doublecortin (DCX) and microtubule-associated protein 2 (MAP2), BPA adversely affected neuronal maturation in neural progenitor cells at concentrations as low as 100 ¥ìM, at which the three traditional methods failed to detect any neurotoxic effect. Our DCX/MAP2 ICC findings indicate that low concentrations of BPA are toxic to developing neurons, and suggest that the devised double ICC technique might provide an effective means of assessing neurotoxic effects of environmental toxins at low concentrations.
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KEYWORD
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Bisphenol A, In vitro toxicology method, Immunocytochemistry, Neurotoxicity, Neuronal maturation
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