Neurochemical Research

, Volume 31, Issue 12, pp 1409–1416

Formation and Rapid Export of the Monochlorobimane–Glutathione Conjugate in Cultured Rat Astrocytes

Original Paper

DOI: 10.1007/s11064-006-9192-x

Cite this article as:
Waak, J. & Dringen, R. Neurochem Res (2006) 31: 1409. doi:10.1007/s11064-006-9192-x

Abstract

Monochlorobimane (MCB) is often used to visualize glutathione (GSH) levels in cultured cells, since it is quickly converted to a fluorescent GSH conjugate (GS–MCB). To test for consequences of MCB application on the GSH metabolism of astrocytes, we have studied rat astrocyte-rich primary cultures as model system. MCB caused a concentration dependent rapid decrease in the cellular GSH content. Simultaneously, a transient accumulation of GS–MCB in the cells was observed with a maximal content 5 min after MCB application. The cellular accumulation was followed by a rapid release of GS–MCB into the medium with a maximal initial export rate of 27.9 ± 6.5 nmol h−1 mg protein−1. Transporters of the family of multidrug resistance proteins (Mrps) are likely to be involved in this export, since the Mrp inhibitor MK571 lowered the export rate by 60%. These data demonstrate that, due to its rapid export from astrocytes, GS–MCB is only under well-defined conditions a reliable indicator of the cellular GSH concentration and that MK571 can be used to maintain maximal GS–MCB levels in astrocytes.

Keywords

Export Glutathione Glutathione-S-transferase MCB Multidrug resistance proteins 

Copyright information

© Springer Science+Business Media, LLC 2006

Authors and Affiliations

  1. 1.Center for Biomolecular Interactions BremenUniversity of BremenBremenGermany
  2. 2.School of Psychology, Psychiatry and Psychological MedicineMonash UniversityClaytonAustralia

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