Activation of the antioxidant response element in primary cortical neuronal cultures derived from transgenic reporter mice

Johnson, Dale G.; Andrews, Glen K.; Xu, Weihua; Johnson, Joy L.

doi:10.1046/j.1471-4159.2002.00913.x

Cited by 145 publications

(171 citation statements)

References 28 publications

(51 reference statements)

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“…To test this idea, we used a reporter construct carrying a heat-stable hPAP gene driven by a minimal AREbearing promoter (rQR51). Previous studies in our lab show that, in rQR51-transfected cortical cultures and cultures derived from transgenic ARE-hPAP reporter mice, hPAP staining is restricted mainly to the astrocyte population Johnson et al, 2002). Consistent with our hypothesis, when rQR51 was cotransfected with an expression vector for Nrf2, the number of neurons staining for hPAP increased dramatically (Fig.…”

Section: Neurons Express Lower Levels Of Nrf2 Protein Than Astrocytessupporting

confidence: 90%

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Coordinate Regulation of Glutathione Biosynthesis and Release by Nrf2-Expressing Glia Potently Protects Neurons from Oxidative Stress

Johnson²,

et al. 2003

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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 (Ͻ 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, ␥-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.

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Section: Neurons Express Lower Levels Of Nrf2 Protein Than Astrocytessupporting

confidence: 90%

“…We previously demonstrated that cortical astrocytes have a higher basal and stimulated level of ARE-mediated gene expression than neurons Johnson et al, 2002). One explanation for this observation would be that astrocytes express higher levels of the transcription factor Nrf2 than neurons.…”

Section: Neurons Express Lower Levels Of Nrf2 Protein Than Astrocytesmentioning

confidence: 93%

Coordinate Regulation of Glutathione Biosynthesis and Release by Nrf2-Expressing Glia Potently Protects Neurons from Oxidative Stress

Johnson²,

et al. 2003

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“…Activation characteristics of the Nrf2-ARE system in SOD models of ALS By using a transgenic mouse line developed in the lab expressing a thermostable human placental alkaline phosphatase (hPAP) reporter construct upon Nrf2 binding to the ARE (AREhPAP mice) [(Johnson et al 2002)], we were able to dissect the activation characteristics of the Nrf2-ARE system throughout the timecourse of ALS pathology in SOD G93A mice. The presence of the ARE-hPAP transgene had no effect on the timecourse of pathology in the mutant SOD animals, as evaluated by time to paralysis, staining for markers of gliosis such as GFAP, and motor neuron cell loss (data not shown).…”

Section: Resultsmentioning

confidence: 99%

“…Mice which express wild-type human Cu/Zn SOD at levels comparable to those seen in the SOD G93A mice (SOD WT mice) serve as a control for elevated levels of human SOD [(Dal Canto and Gurney 1995)]. ARE-hPAP mice containing the core ARE sequence from the rat NADPH: quinone oxidoreductase 1 (NQO1) gene that drives a thermostable hPAP were previously developed and characterized [(Johnson et al 2002)]. Mice were maintained on a heterozygous background so that each litter would generate littermate controls.…”

Section: Animalsmentioning

confidence: 99%

Activation of the Nrf2–ARE pathway in muscle and spinal cord during ALS-like pathology in mice expressing mutant SOD1

Kraft

Resch

Johnson

et al. 2007

Experimental Neurology

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Oxidative stress plays a key role in the neuronal loss exhibited in amyotrophic lateral sclerosis (ALS), an event precipitating irreversible muscle atrophy. By crossing ALS mouse models (SOD G93A and SOD H46RH48Q ) with an antioxidant response element (ARE) reporter mouse, we identified activation characteristics of the ARE system throughout the timecourse of motor neuron disease. Surprisingly, the earliest and most significant activation of this genetic sensor of oxidative stress occurred in the distal muscles of mutant SOD mice. The resultant data supports existing hypotheses that the muscle is somehow implicated during the initial pathology of these mice. Subsequently, Nrf2-ARE activation appears to progress in a retrograde fashion along the motor pathway. These data provide timely information concerning the contributions of the Nrf2-ARE pathway in ALS disease progression.

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“…In addition, it has been reported recently that the PI 3-kinase inhibitors wortmannin and LY-294002 inhibit nuclear translocation of Nrf2 and ARE-mediated transactivation induced by tBHQ (25,27). Hence, it is possible that aPKC transmit signals originating from PI 3-kinase to cause nuclear translocation of Nrf2.…”

Section: Discussionmentioning

confidence: 99%

Atypical protein kinase C mediates activation of NF-E2-related factor 2 in response to oxidative stress

Numazawa

Ishikawa

Yoshida

et al. 2003

American Journal of Physiology-Cell Physiology

305

189

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Transcription factor NF-E2-related factor 2 (Nrf2) regulates the induction of antioxidative proteins, including heme oxygenase-1 (HO-1). Nrf2 is sequestered in the cytoplasm by Keap1 under unstimulated conditions but translocates into the nucleus and transactivates the antioxidant responsive element (ARE) upon exposure to oxidative insults. It has recently been demonstrated that in vitro phosphorylation of Nrf2 on Ser40 by protein kinase C (PKC) facilitates the dissociation of Nrf2 from the Keap1 complex (Huang HC, Nguyen T, and Pickett CB. J Biol Chem 277: 42769–42774, 2002). The present study was designed to examine whether PKC is involved in oxidative stress-mediated nuclear translocation of Nrf2 in vivo and, if so, which PKC isoforms are involved. Induction of HO-1 gene expression by phorone, a glutathione depletor, and 4-hydroxy-2,3-nonenal (4-HNE), an end product of lipid peroxidation, was suppressed by a specific PKC inhibitor, Ro-31-8220, at concentrations that inhibit all isoforms in WI-38 cells. The induction of HO-1 was not affected by prolonged exposure of the cells to 12- O-tetradecanoylphorbol-13 acetate (TPA), suggesting that TPA-insensitive atypical PKC (aPKC) isoforms are involved. An immunocomplex kinase assay revealed that phorone and 4-HNE increased aPKCι activity. In COS-7 cells, 4-HNE induced nuclear translocation of the Nrf2-green fluorescent protein (GFP) fusion protein, but not the Nrf2(S40A)-GFP mutant. In the absence of oxidative insults, the Nrf2(S40E)-GFP mutant was distributed in the nucleus. The Nrf2-GFP accumulation in the nucleus was induced by coexpression of aPKCι, but not by a kinase inactive mutant aPKCι(K274W). The activity of an ARE-driven reporter was increased by coexpression of aPKCι, and this effect was eliminated by Ro-31-8220 in HepG2 cells. The reporter activity induced by 4-HNE was inhibited by coexpression of aPKCι(K274W). These results suggest that phosphorylation of Nrf2 Ser40 by aPKC(s) is involved in the nuclear translocation and ARE transactivation of Nrf2 by oxidative stress.

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Activation of the antioxidant response element in primary cortical neuronal cultures derived from transgenic reporter mice

Cited by 145 publications

References 28 publications

Coordinate Regulation of Glutathione Biosynthesis and Release by Nrf2-Expressing Glia Potently Protects Neurons from Oxidative Stress

Coordinate Regulation of Glutathione Biosynthesis and Release by Nrf2-Expressing Glia Potently Protects Neurons from Oxidative Stress

Activation of the Nrf2–ARE pathway in muscle and spinal cord during ALS-like pathology in mice expressing mutant SOD1

Atypical protein kinase C mediates activation of NF-E2-related factor 2 in response to oxidative stress

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