Comparative sensitivity of rat cerebellar neurons to dysregulation of divalent cation homeostasis and cytotoxicity caused by methylmercury.
The objective of the present study was to determine the relative effectiveness of methylmercury (MeHg) to alter divalent cation homeostasis and cause cell death in MeHg-resistant cerebellar Purkinje and MeHg-sensitive granule neurons. Application of 0.5-5 microM MeHg to Purkinje and granule cells grown in culture caused a concentration- and time-dependent biphasic increase in fura-2 fluorescence. At 0.5 and 1 microM MeHg, the elevations of fura-2 fluorescence induced by MeHg were biphasic in both cell types, but significantly delayed in Purkinje as compared to granule cells. Application of the heavy-metal chelator, TPEN, to Purkinje cells caused a precipitous decline in a proportion of the fura-2 fluorescence signal, indicating that MeHg causes release of Ca(2+) and non-Ca(2+) divalent cations. Purkinje cells were also more resistant than granule cells to the neurotoxic effects of MeHg. At 24.5 h after-application of 5 microM MeHg, 97.7% of Purkinje cells were viable. At 3 microM MeHg there was no detectable loss of Purkinje cell viability. In contrast, only 40.6% of cerebellar granule cells were alive 24.5 h after application of 3 microM MeHg. In conclusion, Purkinje neurons in primary cultures appear to be more resistant to MeHg-induced dysregulation of divalent cation homeostasis and subsequent cell death when compared to cerebellar granule cells. There is a significant component of non-Ca(2+) divalent cation released by MeHg in Purkinje neurons.
Edwards
Joshua R
JR
Department of Pharmacology and Toxicology, Michigan State University, B331 Life Sciences Building, East Lansing, MI 48824-1317, USA.
Marty
M Sue
MS
Atchison
William D
WD
D000393:Air Pollutants / Q000737:chemistry / Q000633:toxicity; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002413:Cations, Divalent / Q000378:metabolism* / Q000633:toxicity; D018929:Cell Culture Techniques / Q000379:methods; D016923:Cell Death / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects*; D002478:Cells, Cultured; D002531:Cerebellum / Q000187:drug effects / Q000473:pathology* / Q000503:physiopathology; D002614:Chelating Agents / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D005029:Ethylenediamines / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D005333:Fetus / Q000473:pathology; D016257:Fura-2 / Q000494:pharmacology; D006706:Homeostasis / Q000187:drug effects / Q000502:physiology*; D008767:Methylmercury Compounds / Q000633:toxicity*; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods; D016202:N-Methylaspartate / Q000494:pharmacology; D009474:Neurons / Q000187:drug effects* / Q000473:pathology / Q000648:ultrastructure; D011689:Purkinje Cells / Q000473:pathology / Q000502:physiology / Q000648:ultrastructure; D051381:Rats
D000393:Air Pollutants; D002413:Cations, Divalent; D002614:Chelating Agents; D005029:Ethylenediamines; D008767:Methylmercury Compounds; D016202:N-Methylaspartate; C044387:N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine; D002118:Calcium; D016257:Fura-2