Expression and functions of glutamate and γ‑aminobutyric acid transporters in ischemic models

Qian,Zhongrun; Lin,Yingying; Xing,Jin; Qiu,Yongming; Ren,Li

doi:10.3892/mmr.2018.8888

Expression and functions of glutamate and γ‑aminobutyric acid transporters in ischemic models

Authors:
- Zhongrun Qian
- Yingying Lin
- Jin Xing
- Yongming Qiu
- Li Ren
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Affiliations: Department of Neurosurgery, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, P.R. China, Department of Neurosurgery, Shanghai Pudong Hospital, Fudan University Pudong Medical Center, Shanghai 201399, P.R. China
Published online on: April 16, 2018 https://doi.org/10.3892/mmr.2018.8888
Pages: 8196-8202
Copyright: © Qian et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Glutamate and γ‑aminobutyric acid (GABA) transporters serve central roles in normal neuronal activity and are associated with numerous pathological brain conditions, including ischemia and epilepsy. However, the interplay between these transporters in ischemia remains unclear. In the present study, the expression levels of the excitatory amino acid carrier 1 (EAAC1) and GABA transporter 1 (GAT1) were analyzed in vivo and in vitro within ischemic models by immunofluorescence, western blot and RT‑qPCR. Cell survival rates were analyzed following altered expression of these transporters within neuronal cells by flow cytometry. Expression levels of EAAC1 were reduced within the cerebrum of focal cerebral ischemic middle cerebral artery occlusion rat models as well as in primary neurons cultured under hypoxia. However, GAT1 expression levels were slightly elevated under ischemic conditions. The altered expression levels of EAAC1 and GAT1 were combined within neuron cells and the effects were investigated. Apoptotic analysis revealed that EAAC1 suppression and overexpression of GAT1 increased neuronal cell apoptosis under hypoxic conditions; however, EAAC1 overexpression combined with GAT1 knockdown reduced neuronal cell apoptosis under hypoxic conditions. The present study detected the expression levels of the glutamate and GABA transporters under hypoxia, in association with ischemia. The results indicated that, increased expression of EAAC1 combined with GAT1 suppression may provide protective effects in the treatment of epilepsy and ischemia.

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1	Choi DW and Rothman SM: The role of glutamate neurotoxicity in hypoxic-ischemic neuronal death. Ann Rev Neurosci. 13:171–182. 1990. View Article : Google Scholar : PubMed/NCBI
2	Mwaniki MK, Atieno M, Lawn JE and Newton CR: Long-term neurodevelopmental outcomes after intrauterine and neonatal insults: A systematic review. Lancet. 379:445–452. 2012. View Article : Google Scholar : PubMed/NCBI
3	Douglas-Escobar M and Weiss MD: Hypoxic-ischemic encephalopathy: A review for the clinician. JAMA Pediatr. 169:397–403. 2015. View Article : Google Scholar : PubMed/NCBI
4	Matsumoto N, Kumamoto E, Furue H and Yoshimura M: GABA-mediated inhibition of glutamate release during ischemia in substantia gelatinosa of the adult rat. J Neurophysiol. 89:257–264. 2003. View Article : Google Scholar : PubMed/NCBI
5	Voytenko LP, Lushnikova IV, Savotchenko AV, Isaeva EV, Skok MV, Lykhmus OY, Patseva MA and Skibo GG: Hippocampal GABAergic interneurons coexpressing alpha7-nicotinic receptors and connexin-36 are able to improve neuronal viability under oxygen-glucose deprivation. Brain Res. 1616:134–145. 2015. View Article : Google Scholar : PubMed/NCBI
6	Zhou Y and Danbolt NC: GABA and glutamate transporters in brain. Front Endocrinol (Lausanne). 4:1652013.PubMed/NCBI
7	Danbolt NC: Glutamate uptake. Prog Neurobiol. 65:1–105. 2001. View Article : Google Scholar : PubMed/NCBI
8	Tanaka K, Watase K, Manabe T, Yamada K, Watanabe M, Takahashi K, Iwama H, Nishikawa T, Ichihara N, Kikuchi T, et al: Epilepsy and exacerbation of brain injury in mice lacking the glutamate transporter GLT-1. Science. 276:1699–1702. 1997. View Article : Google Scholar : PubMed/NCBI
9	Aoyama K, Suh SW, Hamby AM, Liu J, Chan WY, Chen Y and Swanson RA: Neuronal glutathione deficiency and age-dependent neurodegeneration in the EAAC1 deficient mouse. Nat Neurosci. 9:119–126. 2006. View Article : Google Scholar : PubMed/NCBI
10	Kiryu-Seo S, Gamo K, Tachibana T, Tanaka K and Kiyama H: Unique anti-apoptotic activity of EAAC1 in injured motor neurons. EMBO J. 25:3411–3421. 2006. View Article : Google Scholar : PubMed/NCBI
11	Conti F, Minelli A and Melone M: GABA transporters in the mammalian cerebral cortex: Localization, development and pathological implications. Brain Res Brain Res Rev. 45:196–212. 2004. View Article : Google Scholar : PubMed/NCBI
12	Pozdnyakova N, Yatsenko L, Parkhomenko N and Himmelreich N: Perinatal hypoxia induces a long-lasting increase in unstimulated gaba release in rat brain cortex and hippocampus. The protective effect of pyruvate. Neurochem Int. 58:14–21. 2011. View Article : Google Scholar : PubMed/NCBI
13	Richards DA and Bowery NG: Comparative effects of the GABA uptake inhibitors, tiagabine and NNC-711, on extracellular GABA levels in the rat ventrolateral thalamus. Neurochem Res. 21:135–140. 1996. View Article : Google Scholar : PubMed/NCBI
14	Richerson GB and Wu Y: Role of the GABA transporter in epilepsy. Adv Exp Med Biol. 548:76–91. 2004. View Article : Google Scholar : PubMed/NCBI
15	Smith MD, Saunders GW, Clausen RP, Frølund B, Krogsgaard-Larsen P, Larsson OM, Schousboe A, Wilcox KS and White HS: Inhibition of the betaine-GABA transporter (mGAT2/BGT-1) modulates spontaneous electrographic bursting in the medial entorhinal cortex (mEC). Epilepsy Res. 79:6–13. 2008. View Article : Google Scholar : PubMed/NCBI
16	Frahm C, Siegel G, Grass S and Witte OW: Stable expression of the vesicular GABA transporter following photothrombotic infarct in rat brain. Neuroscience. 140:865–877. 2006. View Article : Google Scholar : PubMed/NCBI
17	Nakajima H, Kubo T, Semi Y, Itakura M, Kuwamura M, Izawa T, Azuma YT and Takeuchi T: A rapid, targeted, neuron-selective, in vivo knockdown following a single intracerebroventricular injection of a novel chemically modified siRNA in the adult rat brain. J Biotechnol. 157:326–333. 2012. View Article : Google Scholar : PubMed/NCBI
18	Guo L, Lan J, Lin Y, Guo P, Nie Q, Mao Q, Ren L and Qiu Y: Hypoxia/ischemia up-regulates Id2 expression in neuronal cells in vivo and in vitro. Neurosci Lett. 554:88–93. 2013. View Article : Google Scholar : PubMed/NCBI
19	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
20	Graham EM, Ruis KA, Hartman AL, Northington FJ and Fox HE: A systematic review of the role of intrapartum hypoxia-ischemia in the causation of neonatal encephalopathy. Am J Obstet Gynecol. 199:587–595. 2008. View Article : Google Scholar : PubMed/NCBI
21	Strasser K, Lueckemann L, Kluever V, Thavaneetharajah S, Hoeber D, Bendix I, Fandrey J, Bertsche A and Felderhoff-Mueser U: Dose-dependent effects of levetiracetam after hypoxia and hypothermia in the neonatal mouse brain. Brain Res. 1646:116–124. 2016. View Article : Google Scholar : PubMed/NCBI
22	Murray DM, Bala P, O'Connor CM, Ryan CA, Connolly S and Boylan GB: The predictive value of early neurological examination in neonatal hypoxic-ischaemic encephalopathy and neurodevelopmental outcome at 24 months. Dev Med Child Neurol. 52:e55–e59. 2010. View Article : Google Scholar : PubMed/NCBI
23	Pettmann B and Henderson CE: Neuronal cell death. Neuron. 20:633–647. 1998. View Article : Google Scholar : PubMed/NCBI
24	Broughton BR, Reutens DC and Sobey CG: Apoptotic mechanisms after cerebral ischemia. Stroke. 40:e331–e339. 2009. View Article : Google Scholar : PubMed/NCBI
25	Fu F, Wu D and Qian C: The MicroRNA-224 inhibitor prevents neuronal apoptosis via targeting spastic paraplegia 7 after cerebral ischemia. J Mol Neurosci. 59:421–429. 2016. View Article : Google Scholar : PubMed/NCBI