title: Effects of prolonged exposure to nanomolar concentrations of methylmercury on voltage-sensitive sodium and calcium currents in PC12 cells.
abstract: The neurotoxicant methylmercury (CH(3)Hg(+)) inhibits voltage-sensitive Na(+) and Ca(2+) currents in neuronal preparations following acute, in vitro, exposure. In the present study, effects on voltage-sensitive Na(+) (I(Na)) and Ca(2+) (I(Ca)) currents in pheochromocytoma (PC12) cells were examined following prolonged exposure to CH(3)Hg(+). When PC12 cells cultured in the presence of nerve growth factor (NGF) for 7 days ('primed') were replated in the presence of NGF and 30 nM CH(3)Hg(+), I(Ca), but not I(Na), amplitude was reduced (29%) significantly approximately 24 h later. Quantitative assessment of morphology indicated that this approximately 24 h exposure to CH(3)Hg(+) significantly reduced neurite length. The N-type voltage-sensitive Ca(2+) channel (VSCC) antagonist omega-conotoxin GVIA (500 pM) was without significant effect on current amplitude or morphology in this exposure protocol. When undifferentiated cells were cultured in the presence of NGF and 10 nM CH(3)Hg(+) for 6 days, I(Ca) and I(Na) amplitude were reduced by 36 and 52%, respectively. I(Ca) at the end of a 150 ms test pulse was also reduced by 40% in CH(3)Hg(+)-treated cells. Thus, both inactivating and non-inactivating I(Ca) were reduced equally. There was no change in [(3)H]saxitoxin or omega-[(125)I]conotoxin GVIA binding, nor were there any morphological alterations in cells treated with CH(3)Hg(+) for 6 days. Omega-conotoxin GVIA (500 pM, 6 days), reduced significantly I(Ca), but not I(Na), but was without effect on morphology. These results demonstrate that prolonged exposure to low concentrations of CH(3)Hg(+) reduces cationic currents in differentiating PC12 cells, but that current reduction is not always associated with morphological alteration.
authors.affiliation: Neurotoxicology Division, National Health and Environmental Effects Research Laboratory, Office of Research and Development, US Environmental Protection Agency, Research Triangle Park, NC 27711, USA. shafer.tim@epa.gov
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Shafer
authors.forename: Connie A
authors.initials: CA
authors.lastname: Meacham
authors.forename: Stanley
authors.initials: S
authors.lastname: Barone
mesh_terms: D000818:Animals; D001667:Binding, Competitive / Q000187:drug effects / Q000502:physiology; D015220:Calcium Channels / Q000187:drug effects* / Q000378:metabolism; D020013:Calcium Signaling / Q000187:drug effects / Q000502:physiology; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D048429:Cell Size / Q000187:drug effects / Q000502:physiology; D002490:Central Nervous System / Q000187:drug effects* / Q000378:metabolism / Q000503:physiopathology; D004305:Dose-Response Relationship, Drug; D004334:Drug Administration Schedule; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D020262:Mercury Poisoning, Nervous System / Q000378:metabolism* / Q000503:physiopathology; D008767:Methylmercury Compounds / Q000633:toxicity*; D008954:Models, Biological; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism; D009498:Neurotoxins / Q000633:toxicity*; D016716:PC12 Cells; D051381:Rats; D012530:Saxitoxin / Q000494:pharmacology; D026941:Sodium Channel Blockers / Q000494:pharmacology; D015222:Sodium Channels / Q000187:drug effects* / Q000378:metabolism; D013779:Tetrodotoxin / Q000494:pharmacology; D020866:omega-Conotoxin GVIA / Q000494:pharmacology
chemical_list: D015220:Calcium Channels; D008767:Methylmercury Compounds; D009498:Neurotoxins; D026941:Sodium Channel Blockers; D015222:Sodium Channels; D012530:Saxitoxin; D013779:Tetrodotoxin; D020866:omega-Conotoxin GVIA