Cell viability and proteomic analysis in cultured neurons exposed to methylmercury

• Published in: Human & experimental toxicology Vol. 26; no. 4; pp. 263 - 272
• Main Authors: Vendrell, Iolanda, Carrascal, Montserrat, Vilaró, Maria-Teresa, Abián, Joaquín, Rodríguez-Farré, Eduard, Suñol, Cristina
• Format: Journal Article Conference Proceeding
• Language: English
• Published: London, England SAGE Publications 01.04.2007; Arnold; Sage Publications Ltd
• Subjects: Animals; Apoptosis; Apoptosis - drug effects; Aspartic Acid - metabolism; Biological and medical sciences; Blotting, Western; Brain; Cell Survival - drug effects; Cells, Cultured; Cellular biology; Chemical and industrial products toxicology. Toxic occupational diseases; Chemical contaminants; Dinoprost - analogs & derivatives; Dinoprost - metabolism; Electrophoresis, Gel, Two-Dimensional; Glutamic Acid - metabolism; Glutamic Acid - physiology; Lipid Peroxidation - drug effects; Mass Spectrometry; Medical sciences; Mercury Poisoning, Nervous System - pathology; Metals and various inorganic compounds; Methylmercury Compounds - toxicity; Mice; Nerve Tissue Proteins - chemistry; Neurons; Neurons - pathology; Oxidative Stress - drug effects; Proteins; Proteomics; Rodents; Synaptic Transmission - physiology; Toxicity; Toxicology; cerebellar granule cells; neurotoxicity; cell proteome; lipid peroxidation; primary cultures; glutamate transport; Cerebellum; Nervous system diseases; Toxicity; Central nervous system; Lipids; Proteome; Glutamate; In vitro; Encephalon; Heavy metal; Proteomics; Excitatory aminoacid; Neurotransmitter; Primary culture; Mercury Organic compounds; Transport; Viability; Peroxidation
• ISSN: 0960-3271; 1477-0903
• DOI: 10.1177/0960327106070455

Methylmercury is an environmental contaminant with special selectivity for cerebellar granule cells. The aim of this study was to determine the effect of long-term methylmercury exposure on cell viability and cellular proteome in cultured cerebellar granule cells. Primary cultures of mice cerebellar granule cells were treated with 0-300 nM methylmercury at 2 days in vitro (div) and afterwards the cells were harvested at 12 div. 100 nM methylmercury produced loss of cell viability, reduced intracellular glutamate content and increased lipid peroxidation. Glutamate transport was not modified by methylmercury treatment. Cell death induced by 300 nM methylmercury at 8 div was apoptotic without producing activation of caspase 3. Extracts of total protein were separated by 2D electrophoresis. Around 800 protein spots were visualized by silver staining in SDS-polyacrylamide gels. Gel images were digitized and protein patterns were analysed by image analysis. Several spots were identified through a combination of peptide mass fingerprinting and matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS). The mitochondrial protein 3-ketoacid-coenzyme A transferase I was decreased up to 39% of controls at concentrations of methylmercury that did not produce cytotoxic effects, whereas the cytoplasmic proteins lactate dehydrogenase chain B and actin did not change. Human & Experimental Toxicology (2007) 26, 263-272