[en] Glutathione (GSH) levels are supposed to determine the vulnerability of many cells towards a wide array of insults. We investigated the effects of chronic inhibition of GSH synthesis and acute depletion of GSH on cerebellar granule neurons in vitro and determined cytoplasmic and mitochondrial GSH with relation to mitochondrial function and generation of reactive oxygen intermediates (ROI). l-buthionine sulfoximine (BSO), which irreversibly blocks gamma-glutamyl-cysteine synthase, led to a time- and concentration-dependent loss of cytoplasmic GSH, while mitochondrial GSH was relatively preserved. No increased generation of ROI was detected over 48 h and the mitochondrial membrane potential was largely maintained. Neuronal degeneration occurred when mitochondrial GSH levels had fallen below 50% of control after 24-36 h. In contrast, direct conjugation of mitochondrial and cytoplasmic GSH with etacrynic acid (EA), resulted in immediate loss of mitochondrial GSH, a large increase of ROI within 2 h, subsequent collapse of the mitochondrial membrane potential and complete cell death within 4-8 h. Electron microscopy studies revealed an as yet unknown change of the chromatin structure to a homogeneous granular pattern after BSO, while EA resulted in typical necrotic changes. No typical features of apoptosis, i.e., no chromatin condensation or DNA fragmentation were detected after GSH depletion after BSO or EA treatment.
Disciplines :
Neurology
Author, co-author :
Wüllner, U; Department of Neurology, Eberhard-Karls-University, Hoppe-Seyler-Str. 3, D-72076, Tübingen, Germany. wuellner@uni-bonn.de
Seyfried, J; Department of Neurology, Eberhard-Karls-Univ., Hoppe-S., Tübingen, Germany
Groscurth, P; Institute of Anatomy, Div. Cell Biol., Univ. Zurich-I., Zürich, Switzerland
Beinroth, S; Department of Neurology, Eberhard-Karls-Univ., Hoppe-S., Tübingen, Germany
Winter, S; Department of Neurology, Eberhard-Karls-Univ., Hoppe-S., Tübingen, Germany
Gleichmann, M; Department of Neurology, Eberhard-Karls-Univ., Hoppe-S., Tübingen, Germany
HENEKA, Michael ; Department of Neurology, Rheinische Friedrich-Wilhelms-U., Bonn, Germany
Löschmann, P.-A.; Department of Neurology, Eberhard-Karls-Univ., Hoppe-S., Tübingen, Germany
Schulz, J B; Department of Neurology, Eberhard-Karls-Univ., Hoppe-S., Tübingen, Germany
Weller, M; Department of Neurology, Eberhard-Karls-Univ., Hoppe-S., Tübingen, Germany
Klockgether, T; Department of Neurology, Rheinische Friedrich-Wilhelms-U., Bonn, Germany
External co-authors :
yes
Language :
English
Title :
Glutathione depletion and neuronal cell death: the role of reactive oxygen intermediates and mitochondrial function.
The excellent technical assistance of I. Müller and L. Dumitrescu is gratefully acknowledged; we thank R. Dringen for helpful discussions. This study was supported by the fortune program of the University of Tübingen (UW) and the BMBF 01 KS 9602.
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