Abstract

Recent in vivo studies suggest that the neurotoxin methylmercury (MeHg) is transported into brain as an L-cysteine complex by amino acid transport system L. To test this hypothesis, the mechanism of MeHg uptake into cultured calf brain capillary endothelial cells, an in vitro model of the blood-brain barrier, was examined. Uptake of Me203Hg-L-cysteine followed Michaelis-Menten kinetics, with a Km of 234 +/- 58 microM (mean +/- S.E.) and a Vmax of 57 +/- 25 pmol.micrograms DNA-1.15 sec-1. Uptake of 10 microM MeHg-L-cysteine was stereoselective and Na+ independent and it was inhibited by the system L substrates L-leucine, 2-amino-2-norbornanecarboxylic acid and L-methionine (5 mM), consistent with transport of MeHg-L-cysteine by the L amino acid carrier. L-Glutamate and methylaminoisobutyric acid, which are transported by the acidic and A amino acid carriers, respectively, had no effect. Moreover, uptake of 3H-L-leucine (5 microM) was inhibited by 1 mM MeHg-L-cysteine is transported into brain capillary endothelial cells by the L carrier. Uptake of other MeHg-thiols was also measured. MeHg-D, L-homocysteine uptake was 82 +/- 11% of MeHg-L-cysteine uptake, whereas uptakes of MeHg complexes of L-penicillamine, dimercaptosuccinic acid, N-acetyl-L-cysteine and glutathione were 57 +/- 16%, 19 +/- 7%, 10 +/- 4% and 8 +/- 5% of MeHg-L-cysteine uptake, respectively. These results illustrate the potential to minimize transport of MeHg across brain capillary endothelium by the careful choice of thiol complexing agent.