The multidrug resistance protein 1 (Mrp1), but not Mrp5, mediates export of glutathione and glutathione disulfide from brain astrocytes.
Astrocytes play an important role in the glutathione (GSH) metabolism of the brain. To test for an involvement of multidrug resistance protein (Mrp) 1 and 5 in the release of GSH and glutathione disulfide (GSSG) from astrocytes, we used astrocyte cultures from wild-type, Mrp1-deficient [Mrp1(-/-)] and Mrp5-deficient [Mrp5(-/-)] mice. During incubation of wild-type or Mrp5(-/-) astrocytes, GSH accumulated in the medium at a rate of about 3 nmol/(h.mg), whereas the export of GSH from Mrp1(-/-) astrocytes was only one-third of that. In addition, Mrp1(-/-) astrocytes had a 50% higher specific GSH content than wild-type or Mrp5(-/-) cells. The presence of 50 microm of the Mrp inhibitor MK571 inhibited the rate of GSH release from wild-type and Mrp5(-/-) astrocytes by 60%, but stimulated at the low concentration of 1 microm GSH release by 40%. In contrast, both concentrations of MK571 did not affect GSH export from Mrp1(-/-) astrocytes. Moreover, in contrast to wild-type and Mrp5(-/-) cells, GSSG export during H(2)O(2) stress was not observed for Mrp1(-/-) astrocytes. These data demonstrate that in astrocytes Mrp1 mediates 60% of the GSH export, that Mrp1 is exclusively responsible for GSSG export and that Mrp5 does not contribute to these transport processes.
Minich
Tobias
T
Institute for Biochemistry, University of Tuebingen, Germany.
Riemer
Jan
J
Schulz
Jörg B
JB
Wielinga
Peter
P
Wijnholds
Jan
J
Dringen
Ralf
R
D000818:Animals; D000831:Animals, Newborn; D001253:Astrocytes / Q000187:drug effects / Q000378:metabolism*; D015152:Blotting, Northern / Q000379:methods; D001921:Brain / Q000166:cytology*; D002470:Cell Survival / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005978:Glutathione / Q000378:metabolism*; D019803:Glutathione Disulfide / Q000378:metabolism*; D006861:Hydrogen Peroxide / Q000494:pharmacology; D020024:Leukotriene Antagonists / Q000494:pharmacology; D051379:Mice; D018345:Mice, Knockout; D015778:Minor Histocompatibility Antigens / Q000494:pharmacology; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000172:deficiency / Q000378:metabolism*; D011422:Propionates / Q000494:pharmacology; D021381:Protein Transport; D011804:Quinolines / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D013997:Time Factors
D004791:Enzyme Inhibitors; D005978:Glutathione; D019803:Glutathione Disulfide; D006861:Hydrogen Peroxide; D020024:Leukotriene Antagonists; D015778:Minor Histocompatibility Antigens; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D011422:Propionates; D011804:Quinolines; D012333:RNA, Messenger; C492376:Abcc5 protein, mouse; C083796:HX antigen; C059141:verlukast; C421109:multidrug resistance-associated protein 1