Purification of Glutathione Reductase from Bovine Brain, Generation of an Antiserum, and Immunocytochemical Localization of the Enzyme in Neural Cells

Gutterer, Jan Mirko; Dringen, Ralf; Hirrlinger, Johannes; Hamprecht, Bernd

doi:10.1046/j.1471-4159.1999.0731422.x

Cited by 69 publications

(35 citation statements)

References 50 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The high specific activities of GPx and GR in microglial cultures are in accordance with the observations that immunoreactivity of GPx in brain slices is most prominent in microglia [Lindenau et al, 1998], and that in astroglia-rich primary cultures, microglial cells stain intensively for GSH and GR [Chatterjee et al, 1999;Gutterer et al, 1999].…”

Section: Discussionsupporting

confidence: 86%

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

et al. 2000

Self Cite

View full text Add to dashboard Cite

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.

show abstract

Section: Discussionsupporting

confidence: 86%

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

et al. 2000

Self Cite

View full text Add to dashboard Cite

show abstract

“…The cultures were used for experiments at an age between 15 and 27 days. Immunocytochemical studies have shown that the astrocyterich cultures comprise more than 90% astrocytes and contain small fractions of oligodendroglial, ependymal, and microglial cells but no neurons (Löffler et al, 1986;Reinhart et al, 1990;Gutterer et al, 1999;Dang et al, 2009). …”

Section: Cell Culturesmentioning

confidence: 99%

Uptake of ferrous iron by cultured rat astrocytes

Tulpule

Robinson

Bishop

et al. 2009

J of Neuroscience Research

Self Cite

View full text Add to dashboard Cite

Astrocytes are considered to play an important role in iron homeostasis of the brain, yet the mechanisms involved in the uptake of iron into astrocytes remain elusive. To investigate the uptake of iron into astrocytes, we have applied ferric ammonium citrate (FAC) to rat astrocyte-rich primary cultures. These cultures express the mRNAs of two membrane-bound ferric reductases, Dcytb and SDR2, and reduce extracellular ferric iron (100 muM) with a rate of 3.2 +/- 0.4 nmol/(hr x mg). This reduction rate is substantially lower than the rate of cellular iron accumulation from 100 muM FAC [24.7 +/- 8.9 nmol/(hr x mg)], which suggests that iron accumulation from FAC does at best partially depend on extracellular ferric reduction. Nonetheless, when the iron in FAC was almost completely reduced by an excess of exogenous ascorbate, astrocytes accumulated iron in a time- and concentration-dependent manner with specific iron accumulation rates that increased linearly for concentrations of up to 100 muM ferrous iron. This accumulation was attenuated by lowering the incubation temperature, by the presence of ferrous iron chelators, or by lowering the pH from 7.4 to 6.8. These data indicate that, in addition to the DMT1-mediated uptake of ferrous iron, astrocytes can accumulate ferric and ferrous iron by mechanisms that are independent of DMT1 or transferrin.

show abstract

“…Although the recycle system of GSSG, comprising GR and NADPH, is the predominant one, few studies have demonstrated the localization of the GR protein. It is present in tissues such as lung cancer cells [10] and brain cells [11,12].…”

Section: Introductionmentioning

confidence: 99%

Alteration of Glutathione Reductase Expression in the Female Reproductive Organs During the Estrous Cycle1

Kaneko¹,

Iuchi²,

Kawachiya³

et al. 2001

Biology of Reproduction

View full text Add to dashboard Cite

The enzyme glutathione reductase (GR) recycles oxidized glutathione (GSSG) by converting it to the reduced form (GSH) in an NADPH-dependent manner. A specific antibody raised against recombinant rat GR was used to localize the protein in the female reproductive organs during the estrous cycle in the rat. In the ovary, the strongest reactivity to the antibody was observed in oocytes, followed by granulosa cells, corpus luteum, and interstitial cells. A strongly positive reaction was also observed mainly in the oviduct epithelia, uterine epithelia, and endometrial gland in the reproductive tract. Oviducts contained the highest GR activity. The GR activity of uterus during metestrus was about twice as high as that for other stages of the cycle. The levels of GR proteins in the tissues roughly matched the activities. The expression of the GR mRNA was highest during metestrus. Because GSH is known to increase gamete viability and the efficiency of fertility, GR, which is expressed in these tissues, is predicted to play a pivotal role in the reproduction process as a source of GSH.

show abstract

Purification of Glutathione Reductase from Bovine Brain, Generation of an Antiserum, and Immunocytochemical Localization of the Enzyme in Neural Cells

Cited by 69 publications

References 50 publications

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

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

Uptake of ferrous iron by cultured rat astrocytes

Alteration of Glutathione Reductase Expression in the Female Reproductive Organs During the Estrous Cycle1

Contact Info

Product

Resources

About