Mechanism of methylmercury efflux from cultured astrocytes.
To study the mechanism of methylmercury (MeHg) efflux from the central nervous system cells, cultured astroglia obtained from neonatal rats were incubated with 10 microM MeHg-cysteine (CySH) for 30 min. After being washed four times, cells were incubated in Hg-free medium, and the release of MeHg from the cells was monitored. The amount of MeHg released in the medium approached a plateau level (ca. 31% of the loaded amount) at 4 hr. Treatment of the cells with a CySH precursor, 2-oxothiazolidine-4-carboxylic acid (OTC), resulted in a significant increase of cellular levels of CySH and glutathione (GSH). OTC also increased 1.5-fold the MeHg efflux from the loaded cells. Another GSH enhancer, GSH isopropyl ester, also stimulated MeHg export from the cells. Ion-exchange column chromatography using DEAE-Sephadex revealed that the MeHg metabolite thus released was exclusively MeHg-GSH conjugate, both with and without OTC. Since the MeHg efflux was suppressed significantly by the presence of probenecid, the efflux occurred via the probenecid-sensitive organic acid transport system. Even though the cellular GSH levels were depleted drastically by treatment with L-buthionine-(S,R)-sulfoximine (BSO), a considerable level (90% of the control) of Hg efflux was detected. Since neither GSH- nor CySH-MeHg was detected in the culture medium of the BSO-treated cells, GSH depletion may trigger some other secretion system(s) in the cells. These results suggest that conjugation with GSH is the major pathway for MeHg efflux in rat astroglia, and that elevation in the cellular GSH level would possibly be a logical therapy for MeHg poisoning, promoting the accelerated elimination of MeHg from the critical tissues.
Fujiyama
J
J
Third Department of Internal Medicine, Faculty of Medicine, Kagoshima University, Japan.
Hirayama
K
K
Yasutake
A
A
D000818:Animals; D001253:Astrocytes / Q000378:metabolism*; D019328:Buthionine Sulfoximine; D002478:Cells, Cultured; D003545:Cysteine / Q000031:analogs & derivatives / Q000494:pharmacology; D005978:Glutathione / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology; D008717:Methionine Sulfoximine / Q000031:analogs & derivatives / Q000494:pharmacology; D008767:Methylmercury Compounds / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D011339:Probenecid / Q000494:pharmacology; D011761:Pyrrolidonecarboxylic Acid; D051381:Rats; D013844:Thiazoles / Q000494:pharmacology; D053778:Thiazolidines
D008767:Methylmercury Compounds; D013844:Thiazoles; D053778:Thiazolidines; C030589:methylmercury cysteine; D008717:Methionine Sulfoximine; D019328:Buthionine Sulfoximine; C058293:glutathione monoisopropyl ester; D005978:Glutathione; D003545:Cysteine; D011339:Probenecid; D011761:Pyrrolidonecarboxylic Acid; C029536:2-oxothiazolidine-4-carboxylic acid