title: Superoxide produced in the matrix of mitochondria enhances methylmercury toxicity in human neuroblastoma cells.
abstract: The mechanism of intracellular metabolism of methylmercury (MeHg) is not fully known. It has been shown that superoxide (O2(-)), the proximal reactive oxygen species (ROS) generated by mitochondria, is responsible for MeHg demethylation. Here, we investigated the impact of different mitochondrial respiratory inhibitors, namely rotenone and antimycin A, on the O2(-)mediated degradation of MeHg in human neuroblastoma cells SH-K-SN. We also utilized paraquat (PQ) which generates O2(-) in the mitochondrial matrix. We found that the cleavage of the carbon-metal bond in MeHg was highly dependent on the topology of O2(-) production by mitochondria. Both rotenone and PQ, which increase O2(-) in the mitochondrial matrix at a dose-dependent manner, enhanced the conversion of MeHg to inorganic mercury (iHg). Surprisingly, antimycin A, which prompts emission of O2(-) into the intermembrane space, did not have the same effect even though antimycin A induced a dose dependent increase in O2(-) emission. Rotenone and PQ also enhanced the toxicity of sub-toxic doses (0.1 μM) MeHg which correlated with the accumulation of iHg in mitochondria and depletion of mitochondrial protein thiols. Taken together, our results demonstrate that MeHg degradation is mediated by mitochondrial O2(-), specifically within the matrix of mitochondria when O2(-) is in adequate supply. Our results also show that O2(-) amplifies MeHg toxicity specifically through its conversion to iHg and subsequent interaction with protein cysteine thiols (R-SH). The implications of our findings in mercury neurotoxicity are discussed herein.
authors.affiliation: University of Ottawa, Department of Biology, Center for Advanced Research in Environmental Genomics, Ottawa, ON, K1N 6N5, Canada.
authors.forename: Ryan J
authors.initials: RJ
authors.lastname: Mailloux
authors.affiliation: University of Ottawa, Department of Biology, Center for Advanced Research in Environmental Genomics, Ottawa, ON, K1N 6N5, Canada.
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Yumvihoze
authors.affiliation: University of Ottawa, Department of Biology, Center for Advanced Research in Environmental Genomics, Ottawa, ON, K1N 6N5, Canada. Electronic address: laurie.chan@uottawa.ca.
authors.forename: Hing Man
authors.initials: HM
authors.lastname: Chan
mesh_terms: D000968:Antimycin A / Q000494:pharmacology; D045744:Cell Line, Tumor; D003545:Cysteine / Q000378:metabolism; D006801:Humans; D008628:Mercury / Q000378:metabolism; D008767:Methylmercury Compounds / Q000378:metabolism*; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism*; D009447:Neuroblastoma / Q000378:metabolism*; D010269:Paraquat / Q000494:pharmacology; D012402:Rotenone / Q000494:pharmacology; D013438:Sulfhydryl Compounds / Q000378:metabolism; D013481:Superoxides / Q000378:metabolism*
chemical_list: D008767:Methylmercury Compounds; D013438:Sulfhydryl Compounds; D012402:Rotenone; D013481:Superoxides; D000968:Antimycin A; D008628:Mercury; D003545:Cysteine; D010269:Paraquat
keywords: Inorganic mercury; Methylmercury; Mitochondria; Neurotoxicity; ROS; Superoxide