Exacerbated vulnerability to oxidative stress in astrocytic C6 glioma cells with stable overexpression of the glutamine transporter slc38a1

Ogura, Masato; Takarada, Takeshi; Nakamichi, Noritaka; Kawagoe, H; Sako, Aya; Nakazato, Ryota; Yoneda, Yukio

doi:10.1016/j.neuint.2011.01.007

Cited by 19 publications

(14 citation statements)

References 39 publications

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“…These include Fmo2, Hmox1, Ift172, Sepp1, Slc38a1, Sod3, Srxn1 , and Ucp2. Greatly enhanced expression of these genes implied a compensatory reaction against oxidative stress that prevailed in the hippocampus of GWI-rats because, proteins coding some of these genes are directly involved in the regulation of ROS production ( Fmo2 , Mao et al, 2010), oxidative cleavage of pro-oxidant damaging molecule heme ( Hmox1 , Xu et al, 2016), transport of oxygen ( Ift172 , Gorivodsky et al, 2009), extracellular antioxidant activity ( Sepp1, Sod3 , Huang et al, 2015; Davis et al, 2017), modulation of intracellular ROS ( Slc38a1 , Ogura et al, 2011), oxidoreductase activity ( Srxn1 , Zhou et al, 2015), and uncoupling of oxidative phosphorylation to reduce mitochondrial ROS ( Ucp2 , Graw et al, 2015). Furthermore, some of the up-regulated genes also encode proteins associated with NF-kB signaling and Nrf2 pathway ( Psmb5 and Sqstm1 ), inflammation ( Mpo ), neuroprotection ( Cat, Cygb, Dhcr24 , and Prnp ), neurocognitive disorders ( Apc and Ctsb ), Alzheimer’s disease ( Dhcr24, Dnm2 , and Nqo1 ), amyotrophic lateral sclerosis ( Als2 ), hippocampal resting state functional connectivity ( Apoe ), reduced neurogenesis ( Mpo ), anti-apoptotic activity ( Dhcr24 ), axon growth and vesicular trafficking ( Dnm2 ), cellular detoxification and stress response ( Gstk1 and Gstp1 ), DNA repair ( Ercc6 ), and altered presynaptic glutamatergic function ( Slc38a1 ).…”

Section: Discussionmentioning

confidence: 99%

Chronic Oxidative Stress, Mitochondrial Dysfunction, Nrf2 Activation and Inflammation in the Hippocampus Accompany Heightened Systemic Inflammation and Oxidative Stress in an Animal Model of Gulf War Illness

Shetty

Hattiangady

Upadhya

et al. 2017

Front. Mol. Neurosci.

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Memory and mood dysfunction are the key symptoms of Gulf war illness (GWI), a lingering multi-symptom ailment afflicting >200,000 veterans who served in the Persian Gulf War-1. Research probing the source of the disease has demonstrated that concomitant exposures to anti-nerve gas agent pyridostigmine bromide (PB), pesticides, and war-related stress are among the chief causes of GWI. Indeed, exposures to GWI-related chemicals (GWIR-Cs) and mild stress in animal models cause memory and mood impairments alongside reduced neurogenesis and chronic low-level inflammation in the hippocampus. In the current study, we examined whether exposure to GWIR-Cs and stress causes chronic changes in the expression of genes related to increased oxidative stress, mitochondrial dysfunction, and inflammation in the hippocampus. We also investigated whether GWI is linked with chronically increased activation of Nrf2 (a master regulator of antioxidant response) in the hippocampus, and inflammation and enhanced oxidative stress at the systemic level. Adult male rats were exposed daily to low-doses of PB and pesticides (DEET and permethrin), in combination with 5 min of restraint stress for 4 weeks. Analysis of the hippocampus performed 6 months after the exposure revealed increased expression of many genes related to oxidative stress response and/or antioxidant activity (Hmox1, Sepp1, and Srxn1), reactive oxygen species metabolism (Fmo2, Sod2, and Ucp2) and oxygen transport (Ift172 and Slc38a1). Furthermore, multiple genes relevant to mitochondrial respiration (Atp6a1, Cox6a1, Cox7a2L, Ndufs7, Ndufv1, Lhpp, Slc25a10, and Ucp1) and neuroinflammation (Nfkb1, Bcl6, Csf2, IL6, Mapk1, Mapk3, Ngf, N-pac, and Prkaca) were up-regulated, alongside 73–88% reduction in the expression of anti-inflammatory genes IL4 and IL10, and nuclear translocation and increased expression of Nrf2 protein. These hippocampal changes were associated with elevated levels of pro-inflammatory cytokines and chemokines (Tnfa, IL1b, IL1a, Tgfb, and Fgf2) and lipid peroxidation byproduct malondialdehyde in the serum, suggesting the presence of an incessant systemic inflammation and elevated oxidative stress. These results imply that chronic oxidative stress, inflammation, and mitochondrial dysfunction in the hippocampus, and heightened systemic inflammation and oxidative stress likely underlie the persistent memory and mood dysfunction observed in GWI.

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Section: Discussionmentioning

confidence: 99%

Chronic Oxidative Stress, Mitochondrial Dysfunction, Nrf2 Activation and Inflammation in the Hippocampus Accompany Heightened Systemic Inflammation and Oxidative Stress in an Animal Model of Gulf War Illness

Shetty

Hattiangady

Upadhya

et al. 2017

Front. Mol. Neurosci.

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“…This product was inserted into pSI, which is a vector for gene transfection, by using the Ligation High regent as described previously [32]. Mouse embryonal carcinoma P19 cells were plated at a density of 1.5×10 5 cells/cm 2 , followed by culture in DMEM with 10% FBS for 24 h and subsequent stable transfection with pSI- GlnT, pSI-GFP and pcDNA3.1 vectors, or pSI, pSI-GFP and pcDNA3.1 vectors, using 2 µg of DNA and Lipofectamine and Plus reagent in 0.5 ml of Opti-MEM described previously [33].…”

Section: Methodsmentioning

confidence: 99%

“…GlnT cDNA including entire coding region was prepared from rat cortical neuronal mRNA by RT-PCR as described previously [32]. cDNA was then subcloned into pT7 blue vector.…”

Section: Methodsmentioning

confidence: 99%

“…An aliquot of supernatants was neutralized with 1 M KHCO 3 , followed by centrifugation and subsequent reaction for 2 min at room temperature with a derivatization reagent [5 mg/ml o-phthalaldehyde, 1% β-mercaptoethanol and 0.36 M potassium borate (pH 10.4)] for HPLC analysis as described previously [32]. An aliquot was injected into an analysis column (Symmetry C 18 3.5 µm, 4.6×75 mm).…”

Section: Methodsmentioning

confidence: 99%

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Promotion of Both Proliferation and Neuronal Differentiation in Pluripotent P19 Cells with Stable Overexpression of the Glutamine Transporter slc38a1

et al. 2012

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BackgroundWe previously demonstrated the functional expression in newborn rat neocortical astrocytes of glutamine transporter (GlnT = slc38a1) believed to predominate in neurons over astroglia in the brain. In order to evaluate the possible role of this transporter in neurogenesis, we attempted to establish stable transfectants of GlnT in mouse embryonal carcinoma P19 cells endowed to proliferate for self-renewal and differentiate into progeny cells such as neurons and astroglia, in addition to in vitro pharmacological profiling of the green tea ingredient theanine, which is shown to be a potent inhibitor of glutamine transport mediated by GlnT in cultured neurons and astroglia.Methodology/Principal FindingsThe full-length coding region of rat GlnT was inserted into a vector for gene transfection along with selection by G418, followed by culture with all-trans retinoic acid under floating conditions and subsequent dispersion for spontaneous differentiation under adherent conditions. Stable overexpression of GlnT led to marked increases in the size of round spheres formed during the culture for 4 days and 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide reduction, with concomitant promotion of subsequent differentiation into cells immunoreactive for a neuronal marker protein. In these stable GlnT transfectants before differentiation, drastic upregulation was seen for mRNA expression of several proneural genes with a basic helix-loop-helix domain such as NeuroD1. Although a drastic increase was seen in NeuroD1 promoter activity in stable GlnT transfectants, theanine doubled NeuroD1 promoter activity in stable transfectants of empty vector (EV), without affecting the promoter activity already elevated in GlnT transfectants. Similarly, theanine promoted cellular proliferation and neuronal differentiation in stable EV transfectants, but failed to further stimulate the acceleration of both proliferation and neuronal differentiation found in stable GlnT transfectants.Conclusions/SignificanceGlnT would promote both proliferation and neuronal differentiation through a mechanism relevant to the upregulation of particular proneural genes in undifferentiated P19 cells.

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“…Overexpression of SNAT was observed in gastric cancer [58], human hepatocellular carcinoma [59], breast cancer [60,61], hilar cholangiocarcinoma [62], C6 glioma [63], prostate cancer [25], HeLa epithelial cervical cancer cells, and 143B osteosarcoma cells [64]; and SNAT expression in stomach and breast cancers is also related to Ki-67 [61] and PCNA (proliferating cell nuclear antigen) expression [58] as indicated by proliferative activity.…”

Section: Trans-1-amino-3-[ 18 F]fluorocyclobutanecarboxylic Acid ([ 1mentioning

confidence: 99%

Relationship between [ 14 C]MeAIB uptake and amino acid transporter family gene expression levels or proliferative activity in a pilot study in human carcinoma cells: Comparison with [ 3 H]methionine uptake

Kagawa

Nishii

Higashi³

et al. 2017

Nuclear Medicine and Biology

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Results and Conclusion: [ 14 C]MeAIB uptake occurred principally via a Na + -dependent A type mechanism whereas [ 3 H]MET uptake, which occurred predominantly via a Na + -independent L type mechanism although other transporters were also utilized depending on cell type. There was no correlation between [ 3 H]MET uptake and total system L amino acid transporter (LAT) expression. In contrast, [ 14 C]MeAIB uptake strongly correlated with total system A amino acid transporter (SNAT) expression and proliferative activity in this preliminary study using four human carcinoma cell lines. Carcinoma proliferative activity also correlated with total SNAT expression.

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Exacerbated vulnerability to oxidative stress in astrocytic C6 glioma cells with stable overexpression of the glutamine transporter slc38a1

Cited by 19 publications

References 39 publications

Chronic Oxidative Stress, Mitochondrial Dysfunction, Nrf2 Activation and Inflammation in the Hippocampus Accompany Heightened Systemic Inflammation and Oxidative Stress in an Animal Model of Gulf War Illness

Chronic Oxidative Stress, Mitochondrial Dysfunction, Nrf2 Activation and Inflammation in the Hippocampus Accompany Heightened Systemic Inflammation and Oxidative Stress in an Animal Model of Gulf War Illness

Promotion of Both Proliferation and Neuronal Differentiation in Pluripotent P19 Cells with Stable Overexpression of the Glutamine Transporter slc38a1

Relationship between [ 14 C]MeAIB uptake and amino acid transporter family gene expression levels or proliferative activity in a pilot study in human carcinoma cells: Comparison with [ 3 H]methionine uptake

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