Mitochondrial glutathione and methyl iodide-induced neurotoxicity in primary neural cell cultures.
The status of both cytosolic and mitochondrial glutathione was studied in primary cultured cerebrocortical cells from fetal mice using the selective membrane-solubilizing properties of digitonin and after exposure to the monohalomethane methyl iodide. A correlation was found between cell injury (assessed by lactate dehydrogenase leakage 24 hr after exposure) and early loss of mitochondrial glutathione (2 hr after exposure), while cell death did not appear directly dependent on cytosolic glutathione depletion. The antioxidants BW 755C (3-amino-1-[m-(trifluoromethyl)phenyl]-2-pyrazoline) and DPPD (N,N'-diphenyl-p-phenylenediamine), and the glutathione precursor N-acetyl-L-cysteine were used to modify cellular responses to methyl iodide. Prevention of cell injury by these reagents was obtained only under conditions where at least 50% of the normal level of mitochondrial glutathione was preserved after methyl iodide exposure. Mitochondrial metabolic activity (reduction of 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide, MTT) was affected by exposure to methyl iodide and correlated with mitochondrial glutathione depletion and cytotoxicity. These findings indicate that the mitochondrial glutathione pool and mitochondrial functions may be the most significant intracellular targets of methyl iodide in neural cultures. Moreover, the present work exemplifies the dependence of neural cell viability on the status of mitochondrial functions and suggests that, as in the liver, mitochondrial glutathione is an important component of cellular homeostasis in nervous tissue.
Bonnefoi
M S
MS
Chemical Industry Institute of Toxicology, Research Triangle Park, North Carolina 27709.
D000818:Animals; D002462:Cell Membrane / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology / Q000187:drug effects*; D004396:Coloring Agents / Q000378:metabolism; D003600:Cytosol / Q000187:drug effects* / Q000378:metabolism; D004072:Digitonin / Q000633:toxicity; D005978:Glutathione / Q000378:metabolism / Q000633:toxicity*; D006847:Hydrocarbons, Iodinated / Q000633:toxicity*; D007425:Intracellular Membranes / Q000187:drug effects; D051379:Mice; D008928:Mitochondria / Q000187:drug effects*; D013778:Tetrazolium Salts / Q000378:metabolism; D013844:Thiazoles / Q000378:metabolism
D004396:Coloring Agents; D006847:Hydrocarbons, Iodinated; D013778:Tetrazolium Salts; D013844:Thiazoles; C014055:methyl iodide; C022616:thiazolyl blue; D005978:Glutathione; D004072:Digitonin