title: Memantine, a Low-Affinity NMDA Receptor Antagonist, Protects against Methylmercury-Induced Cytotoxicity of Rat Primary Cultured Cortical Neurons, Involvement of Ca2+Dyshomeostasis Antagonism, and Indirect Antioxidation Effects.
abstract: Methylmercury (MeHg) is an extremely dangerous environmental pollutant that induces severe toxic effects in the central nervous system. Neuronal damage plays critical roles mediating MeHg-induced loss of brain function and neurotoxicity. The molecular mechanisms of MeHg neurotoxicity are incompletely understood. The objective of the study is to explore mechanisms that contribute to MeHg-induced neurocyte injuries focusing on neuronal Ca2+dyshomeostasis and alteration of N-methyl-D-aspartate receptors (NMDARs) expression, as well as oxidative stress in primary cultured cortical neurons. In addition, the neuroprotective effects of memantine against MeHg cytotoxicity were also investigated. The cortical neurons were exposed to 0, 0.01, 0.1, 1, or 2 μM methylmercury chloride (MeHgCl) for 0.5-12 h, or pre-treated with 2.5, 5, 10, or 20 μM memantine for 0.5-6 h, respectively; cell viability and LDH release were then quantified. For further experiments, 2.5, 5, and 10 μM of memantine pre-treatment for 3 h followed by 1 μM MeHgCl for 6 h were performed for evaluation of neuronal injuries, specifically addressing apoptosis; intracellular free Ca2+concentrations; ATPase activities; calpain activities; expressions of NMDAR subunits (NR1, NR2A, NR2B); NPSH levels; and ROS formation. Exposure of MeHgCl resulted in toxicity of cortical neurons, which were shown as a loss of cell viability, high levels of LDH release, morphological changes, and cell apoptosis. Moreover, intracellular Ca2+dyshomeostasis, ATPase activities inhibition, calpain activities, and NMDARs expression alteration were observed with 1 μM MeHgCl administration. Last but not least, NPSH depletion and reactive oxygen species (ROS) overproduction showed an obvious oxidative stress in neurons. However, memantine pre-treatment dose-dependently antagonized MeHg-induced neuronal toxic effects, apoptosis, Ca2+dyshomeostasis, NMDARs expression alteration, and oxidative stress. In conclusion, the cytoprotective effects of memantine against MeHg appeared to be mediated not only via its NMDAR binding properties and Ca2+homeostasis maintenance but also by indirect antioxidation effects.
authors.affiliation: Department of environmental health, school of public health, China Medical University, Shenyang, People's Republic of China.
authors.forename: Wei
authors.initials: W
authors.lastname: Liu
authors.affiliation: Department of environmental health, school of public health, China Medical University, Shenyang, People's Republic of China. zfxu@mail.cmu.edu.cn.
authors.forename: Zhaofa
authors.initials: Z
authors.lastname: Xu
authors.affiliation: Department of environmental health, school of public health, China Medical University, Shenyang, People's Republic of China.
authors.forename: Tianyao
authors.initials: T
authors.lastname: Yang
authors.affiliation: Department of environmental health, school of public health, China Medical University, Shenyang, People's Republic of China.
authors.forename: Bin
authors.initials: B
authors.lastname: Xu
authors.affiliation: Department of environmental health, school of public health, China Medical University, Shenyang, People's Republic of China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Deng
authors.affiliation: Department of environmental health, school of public health, China Medical University, Shenyang, People's Republic of China.
authors.forename: Shu
authors.initials: S
authors.lastname: Feng
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D002478:Cells, Cultured; D005260:Female; D006706:Homeostasis / Q000187:drug effects*; D008297:Male; D008559:Memantine / Q000494:pharmacology*; D008767:Methylmercury Compounds / Q000494:pharmacology; D009474:Neurons / Q000187:drug effects / Q000378:metabolism; D018696:Neuroprotective Agents / Q000494:pharmacology; D018384:Oxidative Stress / Q000187:drug effects*; D017208:Rats, Wistar; D017382:Reactive Oxygen Species / Q000378:metabolism; D016194:Receptors, N-Methyl-D-Aspartate / Q000187:drug effects* / Q000378:metabolism
chemical_list: D000975:Antioxidants; D008767:Methylmercury Compounds; D018696:Neuroprotective Agents; D017382:Reactive Oxygen Species; D016194:Receptors, N-Methyl-D-Aspartate; C004925:methylmercuric chloride; D008559:Memantine
keywords: Ca2+ dyshomeostasis; Excitotoxicity; Memantine; Methylmercury; NMDA receptors; Neurons; Oxidative stress