title: Methylmercury cytotoxicity in PC12 cells is mediated by primary glutathione depletion independent of excess reactive oxygen species generation.
abstract: Low doses, chronic exposure to mercurial organic compounds is a worldwide health concern and could be pathogenetically relevant as co-factor in several neurodegenerative diseases. In this in vitro study we wanted to further improve our knowledge on the mechanisms of toxicity of methylmercury hydroxide (MeHgOH) in the unprimed PC12 cell line. Cell viability, mitochondrial function, redox state, and cell morphology were recorded at different time points to sequence the events leading to cell death. The lowest cytotoxic concentration and EC50 were 0.3 and 1.3 microM, respectively. 5 microM MeHgOH was fatal for 80% of the cell population after 24 h; within 1 h it caused glutathione (GSH) depletion and a partial dissipation of Deltapsim. At this concentration, reactive oxygen species (ROS) generation was only slight and delayed. After 6h more than 50% of ATP was available and caspase 3 was active. Time-lapse confocal microscopy showed that only a fraction of the cells completed apoptosis while others turned toward necrosis (necrapoptosis). Pre-incubation with N-acetylcysteine (NAC) and GSH but not Cyclosporin A rescued over 80% of the cells. These results provide experimental evidence that, in this cell model, MeHgOH triggers cell death via a primary depletion of GSH but in the absence of ROS overproduction.
authors.affiliation: Department of Experimental Medicine, via Volturno 39, Università degli Studi di Parma, 43100 Parma, Italy. rita.gatti@unipr.it
authors.forename: Rita
authors.initials: R
authors.lastname: Gatti
authors.forename: Silvana
authors.initials: S
authors.lastname: Belletti
authors.forename: Jacopo
authors.initials: J
authors.lastname: Uggeri
authors.forename: Maria Vittoria
authors.initials: MV
authors.lastname: Vettori
authors.forename: Antonio
authors.initials: A
authors.lastname: Mutti
authors.forename: Renato
authors.initials: R
authors.lastname: Scandroglio
authors.forename: Guido
authors.initials: G
authors.lastname: Orlandini
mesh_terms: D000818:Animals; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D005978:Glutathione / Q000378:metabolism*; D007424:Intracellular Fluid / Q000187:drug effects / Q000378:metabolism; D008767:Methylmercury Compounds / Q000633:toxicity*; D016716:PC12 Cells; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism*
chemical_list: D008767:Methylmercury Compounds; D017382:Reactive Oxygen Species; D005978:Glutathione