Glutathione depletion modulates methanol, formaldehyde and formate toxicity in cultured rat conceptuses.
The proposed use of methanol (H3COH) as an alternative to fossil fuels has prompted concern about potential health risks resulting from widespread environmental exposure. Methanol is teratogenic in rodents and, although the exact toxic species is not known, teratogenesis may result from the enzymatic biotransformation of H3COH to formaldehyde (CH2O) and formic acid causing increased biological reactivity and toxicity. A protective role for the antioxidant glutathione (GSH) has been described for H3COH, CH2O and formic acid toxicity in various biological systems but has yet to be evaluated in the developing conceptus. Whole embryo culture studies were conducted using GD 10-11 rat conceptuses to elucidate the relationship between H3COH and its metabolites and GSH status. Methanol exposure produced a decrease in normal growth parameters and a dose-dependent loss of viability. CH2O had deleterious effects on embryo growth and viability. Sodium formate (HCOONa) exposure resulted in a high mortality rate but viable embryos did not manifest any abnormalities. Methanol, CH2O, and HCOONa all produced a significant depletion of GSH in both embryo and VYS. Inhibition of GSH synthesis by L-buthionine-S,R-sulfoximine (BSO) treatment exacerbated H3COH, CH2O and HCOONa embryotoxicity. Interestingly, only H3COH/BSO and CH2O/BSO co-treatments caused increased malformation, while embryos treated with HCOONa/BSO did not produce any developmental deformities. These results implicate CH2O as the most embryotoxic H3COH metabolite, on a molar basis, in terms of causing dysmorphogenesis, alterations of normal growth parameters and embryolethality. HCOONa was selectively embryolethal and did not produce dysmorphogenesis. CH2O toxicity is potentiated by GSH depletion, indicating that GSH may be more directly involved in its detoxication in the embryo.
Harris
C
C
Toxicology Program, Department of Environmental Health Sciences, University of Michigan, Ann Arbor, Michigan 48109-2029, USA. charris@umich.edu
Dixon
M
M
Hansen
J M
JM
D000818:Animals; D019328:Buthionine Sulfoximine / Q000494:pharmacology; D003545:Cysteine / Q000096:biosynthesis; D004622:Embryo, Mammalian / Q000187:drug effects* / Q000196:embryology / Q000378:metabolism; D005260:Female; D005557:Formaldehyde / Q000494:pharmacology*; D005561:Formates / Q000494:pharmacology*; D005978:Glutathione / Q000096:biosynthesis*; D066298:In Vitro Techniques; D008297:Male; D000432:Methanol / Q000494:pharmacology*; D011247:Pregnancy; D051381:Rats; D017207:Rats, Sprague-Dawley
D005561:Formates; C030544:formic acid; D005557:Formaldehyde; D019328:Buthionine Sulfoximine; D005978:Glutathione; D003545:Cysteine; D000432:Methanol