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PMID: 12716947 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Coordinate regulation of glutathione biosynthesis and release by Nrf2-expressing glia potently protects neurons from oxidative stress.

The Journal of neuroscience : the official journal of the Society for Neuroscience ·Vol. 23 ·No. 8 ·2003-04-15 ·Pages 3394-406

Shih AY, Johnson DA, Wong G, Kraft AD, Jiang L, Erb H, Johnson JA, Murphy TH

Abstract

Astrocytes have a higher antioxidant potential in comparison to neurons. Pathways associated with this selective advantage include the transcriptional regulation of antioxidant enzymes via the action of the Cap'n'Collar transcription factor Nrf2 at the antioxidant response element (ARE). Here we show that Nrf2 overexpression can reengineer neurons to express this glial pathway and enhance antioxidant gene expression. However, Nrf2-mediated protection from oxidative stress is conferred primarily by glia in mixed cultures. The antioxidant properties of Nrf2-overexpressing glia are more pronounced than those of neurons, and a relatively small number of these glia (< 1% of total cell number added) could protect fully cocultured naive neurons from oxidative glutamate toxicity associated with glutathione (GSH) depletion. Microarray and biochemical analyses indicate a coordinated upregulation of enzymes involved in GSH biosynthesis (xCT cystine antiporter, gamma-glutamylcysteine synthetase, and GSH synthase), use (glutathione S-transferase and glutathione reductase), and export (multidrug resistance protein 1) with Nrf2 overexpression, leading to an increase in both media and intracellular GSH. Selective inhibition of glial GSH synthesis and the supplementation of media GSH indicated that an Nrf2-dependent increase in glial GSH synthesis was both necessary and sufficient for the protection of neurons, respectively. Neuroprotection was not limited to overexpression of Nrf2, because activation of endogenous glial Nrf2 by the small molecule ARE inducer, tert-butylhydroquinone, also protected against oxidative glutamate toxicity.

MeSH Terms
Adenoviridae/genetics Animals Antioxidants/metabolism,pharmacology Apoptosis/drug effects Bridged Bicyclo Compounds/metabolism Cell Death/drug effects Cells, Cultured Coculture Techniques DNA-Binding Proteins/biosynthesis,genetics Enzyme Inhibitors/pharmacology Glutamic Acid/toxicity Glutathione/analogs & derivatives,biosynthesis,metabolism,pharmacology Humans Hydroquinones/pharmacology NF-E2-Related Factor 2 Neuroglia/cytology,metabolism Neurons/cytology,drug effects,metabolism Oligonucleotide Array Sequence Analysis Oxidative Stress/drug effects,physiology Rats Rats, Wistar Response Elements Staurosporine/pharmacology Trans-Activators/biosynthesis,genetics
Chemicals
Antioxidants Bridged Bicyclo Compounds DNA-Binding Proteins Enzyme Inhibitors Hydroquinones NF-E2-Related Factor 2 NFE2L2 protein, human Nfe2l2 protein, rat Trans-Activators glutathione-bimane Glutamic Acid 2-tert-butylhydroquinone Glutathione Staurosporine
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Shih Andy Y
Kinsmen Laboratory of Neurological Research, University of British Columbia, Vancouver, British Columbia, V6T 1Z3, Canada.
Johnson Delinda A
Wong Gloria
Kraft Andrew D
Jiang Lei
Erb Heidi
Johnson Jeffrey A
Murphy Timothy H
References (84)
84 references, click to expand
  1. Activation of antioxidant/electrophile-responsive elements in IMR-32 human neuroblastoma cells.
    Arch Biochem Biophys. 1999 Mar 1;363(1):98-106 PMID: 10049503
  2. Cloning and expression of a plasma membrane cystine/glutamate exchange transporter composed of two distinct proteins.
    J Biol Chem. 1999 Apr 23;274(17):11455-8 PMID: 10206947
  3. GSH transport in immortalized mouse brain endothelial cells: evidence for apical localization of a sodium-dependent GSH transporter.
    J Neurochem. 1999 Jul;73(1):390-9 PMID: 10386992
  4. Evidence of glutathione transporter in rat brain synaptosomal membrane vesicles.
    Neurochem Int. 1999 Jun;34(6):509-16 PMID: 10402226
  5. Nrf2, a Cap'n'Collar transcription factor, regulates induction of the heme oxygenase-1 gene.
    J Biol Chem. 1999 Sep 10;274(37):26071-8 PMID: 10473555
  6. Changes in amino acid concentrations over time and space around an impact injury and their diffusion through the rat spinal cord.
    Exp Neurol. 1999 Oct;159(2):538-44 PMID: 10506525
  7. Regulatory mechanisms of cellular response to oxidative stress.
    Free Radic Res. 1999 Oct;31(4):319-24 PMID: 10517536
  8. Astrocytes prevent neuronal death induced by reactive oxygen and nitrogen species.
    Glia. 1999 Nov;28(2):85-96 PMID: 10533053
  9. Nrf2 is essential for protection against acute pulmonary injury in mice.
    Proc Natl Acad Sci U S A. 1999 Oct 26;96(22):12731-6 PMID: 10535991
  10. GSH transport in human cerebrovascular endothelial cells and human astrocytes: evidence for luminal localization of Na+-dependent GSH transport in HCEC.
    Brain Res. 2000 Jan 10;852(2):374-82 PMID: 10678765
  11. Astrocytes provide cysteine to neurons by releasing glutathione.
    J Neurochem. 2000 Apr;74(4):1434-42 PMID: 10737599
  12. Transcriptional regulation of the antioxidant response element. Activation by Nrf2 and repression by MafK.
    J Biol Chem. 2000 May 19;275(20):15466-73 PMID: 10747902
  13. Evaluation of glutathione-sensitive fluorescent dyes in cortical culture.
    Glia. 2000 Jun;30(4):329-41 PMID: 10797613
  14. The Nrf2 transcription factor contributes both to the basal expression of glutathione S-transferases in mouse liver and to their induction by the chemopreventive synthetic antioxidants, butylated hydroxyanisole and ethoxyquin.
    Biochem Soc Trans. 2000 Feb;28(2):33-41 PMID: 10816095
  15. Transcription factor Nrf2 coordinately regulates a group of oxidative stress-inducible genes in macrophages.
    J Biol Chem. 2000 May 26;275(21):16023-9 PMID: 10821856
  16. Glutathione metabolism in brain metabolic interaction between astrocytes and neurons in the defense against reactive oxygen species.
    Eur J Biochem. 2000 Aug;267(16):4912-6 PMID: 10931173
  17. Thioredoxin reductase and glutathione synthesis is upregulated by t-butylhydroquinone in cortical astrocytes but not in cortical neurons.
    Glia. 2000 Sep;31(3):241-8 PMID: 10941150
  18. Nrf2-dependent activation of the antioxidant responsive element by tert-butylhydroquinone is independent of oxidative stress in IMR-32 human neuroblastoma cells.
    Biochem Biophys Res Commun. 2001 Jan 12;280(1):286-92 PMID: 11162512
  19. The multidrug resistance protein MRP1 mediates the release of glutathione disulfide from rat astrocytes during oxidative stress.
    J Neurochem. 2001 Jan;76(2):627-36 PMID: 11208926
  20. Preferential expression of antioxidant response element mediated gene expression in astrocytes.
    J Neurochem. 2001 Mar;76(6):1670-8 PMID: 11259485
  21. xCt cystine transporter expression in HEK293 cells: pharmacology and localization.
    Biochem Biophys Res Commun. 2001 Apr 20;282(5):1132-7 PMID: 11302733
  22. Astrocytes protect neurons from nitric oxide toxicity by a glutathione-dependent mechanism.
    J Neurochem. 2001 Jun;77(6):1601-10 PMID: 11413243
  23. Oxidative glutamate toxicity can be a component of the excitotoxicity cascade.
    J Neurosci. 2001 Oct 1;21(19):7455-62 PMID: 11567035
  24. Microarray analysis reveals an antioxidant responsive element-driven gene set involved in conferring protection from an oxidative stress-induced apoptosis in IMR-32 cells.
    J Biol Chem. 2002 Jan 4;277(1):388-94 PMID: 11687587
  25. c-Jun mediates axotomy-induced dopamine neuron death in vivo.
    Proc Natl Acad Sci U S A. 2001 Nov 6;98(23):13385-90 PMID: 11687617
  26. Pathways of neuron-astrocyte interactions and their possible role in neuroprotection.
    Eur Arch Psychiatry Clin Neurosci. 2001 Aug;251(4):159-69 PMID: 11697580
  27. Aminopeptidase N mediates the utilization of the GSH precursor CysGly by cultured neurons.
    J Neurosci Res. 2001 Dec 1;66(5):1003-8 PMID: 11746430
  28. Powerful and prolonged protection of human retinal pigment epithelial cells, keratinocytes, and mouse leukemia cells against oxidative damage: the indirect antioxidant effects of sulforaphane.
    Proc Natl Acad Sci U S A. 2001 Dec 18;98(26):15221-6 PMID: 11752465
  29. Role of NRF2 in protection against hyperoxic lung injury in mice.
    Am J Respir Cell Mol Biol. 2002 Feb;26(2):175-82 PMID: 11804867
  30. Site-directed mutagenesis of cysteine to serine in the DNA binding region of Nrf2 decreases its capacity to upregulate antioxidant response element-mediated expression and antioxidant induction of NAD(P)H:quinone oxidoreductase1 gene.
    Oncogene. 2002 Mar 28;21(14):2191-200 PMID: 11948402
  31. Activation of the mouse heme oxygenase-1 gene by 15-deoxy-Delta(12,14)-prostaglandin J(2) is mediated by the stress response elements and transcription factor Nrf2.
    Antioxid Redox Signal. 2002 Apr;4(2):249-57 PMID: 12006176
  32. Time-dependent changes in ARE-driven gene expression by use of a noise-filtering process for microarray data.
    Physiol Genomics. 2002;9(3):137-44 PMID: 12045294
  33. Activation of the antioxidant response element in primary cortical neuronal cultures derived from transgenic reporter mice.
    J Neurochem. 2002 Jun;81(6):1233-41 PMID: 12068071
  34. Glutathione release from cultured brain cells: multidrug resistance protein 1 mediates the release of GSH from rat astroglial cells.
    J Neurosci Res. 2002 Aug 1;69(3):318-26 PMID: 12125073
  35. Drug targeting. Breaking down barriers.
    Science. 2002 Aug 16;297(5584):1116-8 PMID: 12183610
  36. Lack of neurodegeneration in transgenic mice overexpressing mutant amyloid precursor protein is associated with increased levels of transthyretin and the activation of cell survival pathways.
    J Neurosci. 2002 Sep 1;22(17):7380-8 PMID: 12196559
  37. Identification of Nrf2-regulated genes induced by the chemopreventive agent sulforaphane by oligonucleotide microarray.
    Cancer Res. 2002 Sep 15;62(18):5196-203 PMID: 12234984
  38. Electrophile response element-mediated induction of the cystine/glutamate exchange transporter gene expression.
    J Biol Chem. 2002 Nov 22;277(47):44765-71 PMID: 12235164
  39. Nrf2 degradation by the ubiquitin proteasome pathway is inhibited by KIAA0132, the human homolog to INrf2.
    Oncogene. 2002 Oct 3;21(44):6829-34 PMID: 12360409
  40. Degradation of transcription factor Nrf2 via the ubiquitin-proteasome pathway and stabilization by cadmium.
    J Biol Chem. 2003 Jan 24;278(4):2396-402 PMID: 12441344
  41. Increased protein stability as a mechanism that enhances Nrf2-mediated transcriptional activation of the antioxidant response element. Degradation of Nrf2 by the 26 S proteasome.
    J Biol Chem. 2003 Feb 14;278(7):4536-41 PMID: 12446695
  42. The antioxidant responsive element. Activation by oxidative stress and identification of the DNA consensus sequence required for functional activity.
    J Biol Chem. 1991 Jun 25;266(18):11632-9 PMID: 1646813
  43. Glutathione turnover in cultured astrocytes: studies with [15N]glutamate.
    J Neurochem. 1990 Jul;55(1):137-45 PMID: 1972386
  44. Xenobiotic-inducible expression of murine glutathione S-transferase Ya subunit gene is controlled by an electrophile-responsive element.
    Proc Natl Acad Sci U S A. 1990 Aug;87(16):6258-62 PMID: 2166952
  45. Immature cortical neurons are uniquely sensitive to glutamate toxicity by inhibition of cystine uptake.
    FASEB J. 1990 Apr 1;4(6):1624-33 PMID: 2180770
  46. Glutamate toxicity in a neuronal cell line involves inhibition of cystine transport leading to oxidative stress.
    Neuron. 1989 Jun;2(6):1547-58 PMID: 2576375
  47. Glutathione is present in high concentrations in cultured astrocytes but not in cultured neurons.
    Brain Res. 1989 Jul 31;493(2):398-401 PMID: 2765907
  48. Exchange of cystine and glutamate across plasma membrane of human fibroblasts.
    J Biol Chem. 1986 Feb 15;261(5):2256-63 PMID: 2868011
  49. Calcium-dependent and -independent release of glutamate from synaptosomes monitored by continuous fluorometry.
    J Neurochem. 1987 Jul;49(1):50-7 PMID: 2884279
  50. Alteration of amino acid content of cerebrospinal fluid from patients with epilepsy.
    Acta Neurol Scand. 1988 Dec;78(6):473-9 PMID: 3223234
  51. Staurosporine, a potent inhibitor of phospholipid/Ca++dependent protein kinase.
    Biochem Biophys Res Commun. 1986 Mar 13;135(2):397-402 PMID: 3457562
  52. High-efficiency transformation of mammalian cells by plasmid DNA.
    Mol Cell Biol. 1987 Aug;7(8):2745-52 PMID: 3670292
  53. Potent and specific inhibition of glutathione synthesis by buthionine sulfoximine (S-n-butyl homocysteine sulfoximine).
    J Biol Chem. 1979 Aug 25;254(16):7558-60 PMID: 38242
  54. Enzymic method for quantitative determination of nanogram amounts of total and oxidized glutathione: applications to mammalian blood and other tissues.
    Anal Biochem. 1969 Mar;27(3):502-22 PMID: 4388022
  55. Differential expression of heme oxygenase-1 in cultured cortical neurons and astrocytes determined by the aid of a new heme oxygenase antibody. Response to oxidative stress.
    Brain Res Mol Brain Res. 1995 May;30(1):37-47 PMID: 7609642
  56. A detailed analysis of hydrogen peroxide-induced cell death in primary neuronal culture.
    Neuroscience. 1995 Aug;67(4):921-32 PMID: 7675214
  57. Cystine uptake and glutathione level in fetal brain cells in primary culture and in suspension.
    J Neurochem. 1993 Nov;61(5):1667-71 PMID: 7901327
  58. Oxidative stress, glutamate, and neurodegenerative disorders.
    Science. 1993 Oct 29;262(5134):689-95 PMID: 7901908
  59. Oxidative stress induces apoptosis in embryonic cortical neurons.
    J Neurochem. 1994 Jan;62(1):376-9 PMID: 7903353
  60. Vitamin E, ascorbate, glutathione, glutathione disulfide, and enzymes of glutathione metabolism in cultures of chick astrocytes and neurons: evidence that astrocytes play an important role in antioxidative processes in the brain.
    J Neurochem. 1994 Jan;62(1):45-53 PMID: 7903354
  61. Glia: the brain's other cells.
    Science. 1994 Nov 11;266(5187):970-2 PMID: 7973679
  62. Maintenance of neuronal glutathione by glial cells.
    J Neurochem. 1993 Nov;61(5):1672-6 PMID: 8228986
  63. The electrophile counterattack response: protection against neoplasia and toxicity.
    Adv Enzyme Regul. 1993;33:281-96 PMID: 8356913
  64. Evidence for the existence of a sodium-dependent glutathione (GSH) transporter. Expression of bovine brain capillary mRNA and size fractions in Xenopus laevis oocytes and dissociation from gamma-glutamyltranspeptidase and facilitative GSH transporters.
    J Biol Chem. 1996 Apr 19;271(16):9754-8 PMID: 8621654
  65. Glia conditioned medium protects fetal rat midbrain neurones in culture from L-DOPA toxicity.
    Neuroreport. 1996 Jan 31;7(2):441-5 PMID: 8730801
  66. The glutathione S-transferase supergene family: regulation of GST and the contribution of the isoenzymes to cancer chemoprotection and drug resistance.
    Crit Rev Biochem Mol Biol. 1995;30(6):445-600 PMID: 8770536
  67. Glutathione efflux from cultured astrocytes.
    J Neurochem. 1996 May;66(5):1876-81 PMID: 8780013
  68. Astrocytes protect neurons from hydrogen peroxide toxicity.
    J Neurosci. 1996 Apr 15;16(8):2553-62 PMID: 8786431
  69. Antioxidant-inducible genes.
    Adv Pharmacol. 1997;38:293-328 PMID: 8895814
  70. Nrf1 and Nrf2 positively and c-Fos and Fra1 negatively regulate the human antioxidant response element-mediated expression of NAD(P)H:quinone oxidoreductase1 gene.
    Proc Natl Acad Sci U S A. 1996 Dec 10;93(25):14960-5 PMID: 8962164
  71. Different preferences in the utilization of amino acids for glutathione synthesis in cultured neurons and astroglial cells derived from rat brain.
    Neurosci Lett. 1996 Nov 29;219(3):211-4 PMID: 8971817
  72. Postnatal retinal ganglion cells in vitro: protection against reactive oxygen species (ROS)-induced axonal degeneration by cocultured astrocytes.
    Brain Res. 1996 Dec 16;743(1-2):56-62 PMID: 9017230
  73. Construction of adenovirus vectors through Cre-lox recombination.
    J Virol. 1997 Mar;71(3):1842-9 PMID: 9032314
  74. An Nrf2/small Maf heterodimer mediates the induction of phase II detoxifying enzyme genes through antioxidant response elements.
    Biochem Biophys Res Commun. 1997 Jul 18;236(2):313-22 PMID: 9240432
  75. Chemoprevention by inducers of carcinogen detoxication enzymes.
    Environ Health Perspect. 1997 Jun;105 Suppl 4:965-70 PMID: 9255588
  76. Broccoli sprouts: an exceptionally rich source of inducers of enzymes that protect against chemical carcinogens.
    Proc Natl Acad Sci U S A. 1997 Sep 16;94(19):10367-72 PMID: 9294217
  77. Astrocyte-mediated enhancement of neuronal survival is abolished by glutathione deficiency.
    Brain Res. 1997 Oct 3;770(1-2):123-30 PMID: 9372211
  78. Amino acid abnormalities in cerebrospinal fluid of patients with parkinsonism and extrapyramidal disorders.
    Neurology. 1976 May;26(5):489-93 PMID: 944400
  79. Histochemical detection of quinone reductase activity in situ using LY 83583 reduction and oxidation.
    J Neurochem. 1998 May;70(5):2156-64 PMID: 9572303
  80. The regulation of reactive oxygen species production during programmed cell death.
    J Cell Biol. 1998 Jun 15;141(6):1423-32 PMID: 9628898
  81. Activation of endogenous antioxidant defenses in neuronal cells prevents free radical-mediated damage.
    J Neurochem. 1998 Jul;71(1):69-77 PMID: 9648852
  82. Nrf2 and Nrf1 in association with Jun proteins regulate antioxidant response element-mediated expression and coordinated induction of genes encoding detoxifying enzymes.
    Oncogene. 1998 Dec 17;17(24):3145-56 PMID: 9872330
  83. Synthesis of the antioxidant glutathione in neurons: supply by astrocytes of CysGly as precursor for neuronal glutathione.
    J Neurosci. 1999 Jan 15;19(2):562-9 PMID: 9880576
  84. Coordinate regulation of NAD(P)H:quinone oxidoreductase and glutathione-S-transferases in primary cultures of rat neurons and glia: role of the antioxidant/electrophile responsive element.
    Glia. 1999 Jan 15;25(2):131-42 PMID: 9890628
Article Info
Journal
The Journal of neuroscience : the official journal of the Society for Neuroscience
Abbr.
J Neurosci
ISSN
1529-2401
Published
2003-04-15
Pages
3394-406
Language
English
Region
United States
NLM ID
8102140
PMCID
PMC6742304
Subset
IM
Grants
NIEHS NIH HHS · R01 ES010042 · United States
NIEHS NIH HHS · ES10042 · United States
NIEHS NIH HHS · ES08089 · United States
NIEHS NIH HHS · ES09090 · United States
NIEHS NIH HHS · R01 ES008089 · United States
NIEHS NIH HHS · R29 ES008089 · United States
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