Microglial Cells in Culture Express a Prominent Glutathione System for the Defense against Reactive Oxygen Species

• Published in: Developmental neuroscience Vol. 22; no. 5-6; pp. 384 - 392
• Main Authors: Hirrlinger, Johannes, Gutterer, Jan Mirko, Kussmaul, Lothar, Hamprecht, Bernd, Dringen, Ralf
• Format: Journal Article
• Language: English
• Published: Basel, Switzerland S. Karger AG 01.09.2000
• Subjects: Animals; Antigens, Differentiation - biosynthesis; Astrocytes - cytology; Astrocytes - drug effects; Astrocytes - metabolism; Catalase - metabolism; Cells, Cultured; Fluorescent Dyes; Glutathione - biosynthesis; Glutathione Disulfide - biosynthesis; Glutathione Peroxidase - metabolism; Glutathione Reductase - metabolism; Hydrogen Peroxide - metabolism; Hydrogen Peroxide - pharmacology; Hypoxanthine - metabolism; Hypoxanthine - pharmacology; Microglia - cytology; Microglia - drug effects; Microglia - metabolism; Mitochondria, Oxidative Stress and Neurodegeneration; Neurons - cytology; Neurons - drug effects; Neurons - metabolism; Oxidative Stress - drug effects; Phagocytosis - drug effects; Phorbol Esters - pharmacology; Rats; Rats, Wistar; Reactive Oxygen Species - metabolism; Superoxides - metabolism; Xanthine Oxidase - metabolism; Oxidative stress; Hydrogen peroxide; Catalase; Neurons; Astrocytes; Superoxide; Radicals; Microglia; Glutathione
• ISBN: 9783805571043; 3805571046
• ISSN: 0378-5866; 1421-9859
• DOI: 10.1159/000017464

To obtain information on the glutathione metabolism of microglial cells, the content of glutathione and activities of enzymes involved in the defense against peroxides were determined for microglia-rich cultures from rat brain. These cultures contain approximately 90% microglia cells as determined by immunocytochemical staining for glial markers, by the phagocytosis activity of the cells and by the production of superoxide after stimulation of the cells with phorbolester. For these cultures, a glutathione content of 41.2 ± 11.2 nmol/mg protein and a specific activity of glutathione reductase of 15.2 ± 3.2 nmol/(min × mg protein) were determined. These values are significantly higher than those found for astroglial or neuronal cultures. In addition, with 68.7 ± 23.5 nmol/(min × mg protein), the specific activity of glutathione peroxidase in microglial cultures was 78% higher than in cultured neurons. The specific catalase activity of microglial cultures was less than 40% that of astroglial or neuronal cultures. Microglial cultures contain only marginal amounts of oxidized glutathione. However, on application of oxidative stress by incubation of microglial cultures with hydrogen peroxide or with the superoxide-producing hypoxanthine/xanthine oxidase system, cellular glutathione was rapidly oxidized. These results demonstrate that microglial cells have a prominent glutathione system, which is likely to reflect the necessity for self-protection against reactive oxygen species when produced by these or surrounding brain cells.