Cell viability and proteomic analysis in cultured neurons exposed to methylmercury.
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.
Vendrell
Iolanda
I
Department of Neurochemistry, Institut d'Investigacions Biomèdiques de Barcelona, Consejo Superior de Investigaciones Científicas, CSIC - IDIBAPS, Barcelona, Spain.
Carrascal
Montserrat
M
Vilaró
Maria-Teresa
MT
Abián
Joaquín
J
Rodríguez-Farré
Eduard
E
Suñol
Cristina
C
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001224:Aspartic Acid / Q000378:metabolism; D015153:Blotting, Western; D002470:Cell Survival / Q000187:drug effects*; D002478:Cells, Cultured; D015237:Dinoprost / Q000031:analogs & derivatives / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D018698:Glutamic Acid / Q000378:metabolism / Q000502:physiology; D015227:Lipid Peroxidation / Q000187:drug effects; D013058:Mass Spectrometry; D020262:Mercury Poisoning, Nervous System / Q000473:pathology*; D008767:Methylmercury Compounds / Q000633:toxicity*; D051379:Mice; D009419:Nerve Tissue Proteins / Q000737:chemistry; D009474:Neurons / Q000473:pathology*; D018384:Oxidative Stress / Q000187:drug effects; D040901:Proteomics; D009435:Synaptic Transmission / Q000502:physiology
D008767:Methylmercury Compounds; D009419:Nerve Tissue Proteins; C075750:8-epi-prostaglandin F2alpha; D001224:Aspartic Acid; D018698:Glutamic Acid; D015237:Dinoprost