Na+ dependent glutamate transporters (EAAT1, EAAT2, and EAAT3) in primary astrocyte cultures: effect of oxidative stress

Miralles, Vicente J.; Martı́nez-López, Itziar; Zaragozá, Rosa; Borrás, Elisa; García, Carlos J. Gómez; Pallardó, Federico V.; Viña, Juan R.

doi:10.1016/s0006-8993(01)03124-9

Cited by 76 publications

(59 citation statements)

References 35 publications

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“…This expression gradually decreased after 24 h. 10 There were no changes in the expression of EAAT1, EAAT2, and EAAT3 proteins in an oxidative stress model. 20 It has been reported that nociceptive input can cause induction of GLT1 transcription. 9 Our results also showed the downregulation of GT expression concurrent with the appearance of hyperalgesia.…”

Section: Discussionmentioning

confidence: 99%

Comparison of changes in mRNA expression of spinal glutamate transporters following induction of two neuropathic pain models

et al. 2010

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Objectives: It is now suspected that different kinds of neuropathic pain syndromes may have significantly different mechanisms. To date, much effort has been made to investigate the function of glutamate transporters (GTs) after nerve injury. The aim of this study is to compare the changes in GTs' mRNA expression levels between two distinct models of peripheral neuropathic pain: chronic constriction nerve injury (CCI) and spared nerve injury (SNI). Methods: Experiments were performed on animal models of mononeuropathy. Several groups of rats were subjected to behavioral experiments before and 4, 7, and 14 days after the induction of mononeuropathy following the CCI and SNI. Allodynia was assessed by Von Frey filaments, and thermal hyperalgesia was assessed by the paw withdrawal tests. To study molecular experiments, the mRNA expression of (GTs) in CCI and SNI rats, reverse transcription polymerase chain reaction (RT-PCR) were used on days 4 and 14. Results and conclusion: The maximum responses of mechanical allodynia and heat hyperalgesia in two distinct neuropathic pain models were detected on day 14. CCI and SNI induced upregulation of three GTs on day 4, which were followed by GTs downregulation in CCI and downregulation of glutamate aspartate transporter (GLAST) and glutamate transporter (GLT)1 in SNI when examined on day 14. These results indicate that there is an inverse correlation between pain responses and expression of GTs, and also changes in expression of spinal GTs may have a critical function in both the induction and maintenance of neuropathic pain in independent peripheral neuropathic pain models.

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Section: Discussionmentioning

confidence: 99%

Comparison of changes in mRNA expression of spinal glutamate transporters following induction of two neuropathic pain models

et al. 2010

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“…Although the BBB determines the availability, and thereby the brain content, of essential amino acids, astrocytes and neurons participate in maintaining the extracellular concentrations. Astrocytes and neurons have Na ϩ -dependent transport systems capable of transporting neutral and acidic amino acids (8,9,29,42). These various systems are actively involved in regulating the concentrations in ECF and are especially important in the maintenance of low concentrations of neurotransmitter amino acids such as glutamate, aspartate, and glycine.…”

Section: Absence Of Bch Inhibition On Cationic Amino Acid Transport mentioning

confidence: 99%

Na⁺-dependent transport of large neutral amino acids occurs at the abluminal membrane of the blood-brain barrier

O’Kane

Hawkins

2003

American Journal of Physiology-Endocrinology and Metabolism

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Several Na+-dependent carriers of amino acids exist on the abluminal membrane of the blood-brain barrier (BBB). These Na+-dependent carriers are in a position to transfer amino acids from the extracellular fluid of brain to the endothelial cells and thence to the circulation. To date, carriers have been found that may remove nonessential, nitrogen-rich, or acidic (excitatory) amino acids, all of which may be detrimental to brain function. We describe here Na+-dependent transport of large neutral amino acids across the abluminal membrane of the BBB that cannot be ascribed to currently known systems. Fresh brains, from cows killed for food, were used. Microvessels were isolated, and contaminating fragments of basement membranes, astrocyte fragments, and pericytes were removed. Abluminal-enriched membrane fractions from these microvessels were prepared. Transport was Na+ dependent, voltage sensitive, and inhibited by 2-aminobicyclo-(2,2,1)-heptane-2-carboxylic acid, a particular inhibitor of the facilitative large neutral amino acid transporter 1 (LAT1) system. The carrier has a high affinity for leucine ( Km 21 ± 7 μM) and is inhibited by other neutral amino acids, including glutamine, histidine, methionine, phenylalanine, serine, threonine, tryptophan, and tyrosine. Other established neutral amino acids may enter the brain by way of LAT1-type facilitative transport. The presence of a Na+-dependent carrier on the abluminal membrane capable of removing large neutral amino acids, most of which are essential, from brain indicates a more complex situation that has implications for the control of essential amino acid content of brain.

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“…Glutamate uptake assay in cortical neurons [10] Cortical neurons were plated on 24- H] glutamate. After 10 min incubation, the reaction was stopped by washing the cells 3 times with ice-cold PBS.…”

Section: Methodsmentioning

confidence: 99%

Stearic acid protects primary cultured cortical neurons against oxidative stress

Wang

Liang

et al. 2007

Acta Pharmacologica Sinica

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Aim: To observe the effects of stearic acid against oxidative stress in primary cultured cortical neurons. Methods: Cortical neurons were exposed to glutamate, hydrogen peroxide (H 2 O 2 ), or NaN 3 insult in the presence or absence of stearic acid. Cell viability of cortical neurons was determined by MTT assay and LDH release. Endogenous antioxidant enzymes activity[superoxide dismutases (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT)] and lipid peroxidation in cultured cortical neurons were evaluated using commercial kits.5 µmol/L; a noncompetitive inhibitor of proliferator-activated receptor (PPAR)α], bisphenol A diglycidyl ether (BADGE; 100 µmol/L; an antagonist of PPARγ), and cycloheximide (CHX; 30 µmol/L, an inhibitor of protein synthesis) were tested for their effects on the neuroprotection afforded by stearic acid. Western blotting was used to determine the PPARγ protein level in cortical neurons. Results: Stearic acid dose-dependently protected cortical neurons against glutamate or H 2 O 2 injury and increased glutamate uptake in cultured neurons. This protection was concomitant to the inhibition of lipid peroxidation and to the promotion activity of Cu/Zn SOD and CAT in cultured cortical neurons. Its neuroprotective effects were completely blocked by BADGE and CHX. After incubation with H 2 O 2 for 24 h, the expression of the PPARγ protein decreased significantly (P<0.05), and the inhibitory effect of H 2 O 2 on the expression of PPARγ can be attenuated by stearic acid. Conclusion: Stearic acid can protect cortical neurons against oxidative stress by boosting the internal antioxidant enzymes. Its neuroprotective effect may be mainly mediated by the activation of PPARγ and new protein synthesis in cortical neurons.

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Na+ dependent glutamate transporters (EAAT1, EAAT2, and EAAT3) in primary astrocyte cultures: effect of oxidative stress

Cited by 76 publications

References 35 publications

Comparison of changes in mRNA expression of spinal glutamate transporters following induction of two neuropathic pain models

Comparison of changes in mRNA expression of spinal glutamate transporters following induction of two neuropathic pain models

Na⁺-dependent transport of large neutral amino acids occurs at the abluminal membrane of the blood-brain barrier

Stearic acid protects primary cultured cortical neurons against oxidative stress

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Na+ dependent glutamate transporters (EAAT1, EAAT2, and EAAT3) in primary astrocyte cultures: effect of oxidative stress

Cited by 76 publications

References 35 publications

Comparison of changes in mRNA expression of spinal glutamate transporters following induction of two neuropathic pain models

Comparison of changes in mRNA expression of spinal glutamate transporters following induction of two neuropathic pain models

Na+-dependent transport of large neutral amino acids occurs at the abluminal membrane of the blood-brain barrier

Stearic acid protects primary cultured cortical neurons against oxidative stress

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Na⁺-dependent transport of large neutral amino acids occurs at the abluminal membrane of the blood-brain barrier