Astrocytes enhance the tolerance of rat cortical neurons to glutamate excitotoxicity

Zhang,Li‑Nan; Wang,Qi; Xian,Xiao‑Hui; Qi,Jie; Liu,Li‑Zhe; Li,Wen‑Bin

doi:10.3892/mmr.2018.9799

Astrocytes enhance the tolerance of rat cortical neurons to glutamate excitotoxicity

Authors:
- Li‑Nan Zhang
- Qi Wang
- Xiao‑Hui Xian
- Jie Qi
- Li‑Zhe Liu
- Wen‑Bin Li
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Affiliations: Department of Pathophysiology, Hebei Medical University, Shijiazhuang, Hebei 050017, P.R. China
Published online on: December 24, 2018 https://doi.org/10.3892/mmr.2018.9799
Pages: 1521-1528
Copyright: © Zhang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Glutamate excitotoxicity is responsible for neuronal death in acute neurological disorders, including stroke, trauma and neurodegenerative diseases. Astrocytes are the main cells for the removal of glutamate in the synaptic cleft and may affect the tolerance of neurons to the glutamate excitotoxicity. Therefore, the present study aimed to investigate the tolerance of rat cortical neurons to glutamate excitotoxicity in the presence and absence of astrocytes. Rat cortical neurons in the presence or absence of astrocytes were exposed to different concentrations of glutamate (10‑2,000 µM) and 10 µM glycine for different incubation periods. After 24 h, the Cell Counting kit‑8 (CCK‑8) assay was used to measure the cytotoxicity to neurons in the presence or absence of astrocytes. According to the results, in the absence of astrocytes, glutamate induced a concentration‑dependent decrease of neuronal survival rate compared with the control rat cortical neurons, and the neurotoxic half‑maximal inhibitory concentration (IC50) at 15, 30 and 60 min was 364.5, 258.5 and 138.3 µM, respectively. Furthermore, in the presence of astrocytes, glutamate induced a concentration‑dependent decrease of neuronal survival rate compared with the control rat cortical neurons, and the neurotoxic IC50 at 15, 30 and 60 min was 1,935, 932.8 and 789.3 µM, respectively. However, astrocytic toxicity was not observed when the rat cortical astrocytes alone were exposed to different concentrations of glutamate (500, 1,000 and 2,000 µM) for 6, 12 and 24 h. In conclusion, the glutamate‑induced neurotoxic IC50 values at 15, 30 and 60 min were respectively higher in the presence of astrocytes as compared with those in the absence of astrocytes, suggesting that astrocytes can protect against rat cortical neuronal acute damage induced by glutamate.

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1	Lerma J, Herranz AS, Herreras O, Abraira V and Martín del Río R: In vivo determination of extracellular concentration of amino acids in the rat hippocampus. A method based on brain dialysis and computerized analysis. Brain Res. 384:145–155. 1986. View Article : Google Scholar : PubMed/NCBI
2	Baker DA, Xi ZX, Shen H, Swanson CJ and Kalivas PW: The origin and neuronal function of in vivo nonsynaptic glutamate. J Neurosci. 22:9134–9141. 2002. View Article : Google Scholar : PubMed/NCBI
3	Nyitrai G, Kékesi KA and Juhász G: Extracellular level of GABA and Glu: In vivo microdialysis-HPLC measurements. Curr Top Med Chem. 6:935–940. 2006. View Article : Google Scholar : PubMed/NCBI
4	Patneau DK and Mayer ML: Structure-activity relationships for amino acid transmitter candidates acting at N-methyl-D-aspartate and quisqualate receptors. J Neurosci. 10:2385–2399. 1990. View Article : Google Scholar : PubMed/NCBI
5	Levy LM, Attwell D, Hoover F, Ash JF, Bjørås M and Danbolt NC: Inducible expression of the GLT-1 glutamate transporter in a CHO cell line selected for low endogenous glutamate uptake. FEBS Lett. 422:339–342. 1998. View Article : Google Scholar : PubMed/NCBI
6	Kutzing MK, Luo V and Firestein BL: Measurement of synchronous activity by microelectrode arrays uncovers differential effects of sublethal and lethal glutamate concentrations on cortical neurons. Ann Biomed Eng. 39:2252–2262. 2011. View Article : Google Scholar : PubMed/NCBI
7	Zhang J, An S, Hu W, Teng M, Wang X, Qu Y, Liu Y, Yuan Y and Wang D: The neuroprotective properties of hericium erinaceus in glutamate-damaged differentiated PC12 cells and an Alzheimer's disease mouse model. Int J Mol Sci. 17:E18102016. View Article : Google Scholar : PubMed/NCBI
8	Cassano T, Pace L, Bedse G, Lavecchia AM, De Marco F, Gaetani S and Serviddio G: Glutamate and mitochondria: Two prominent players in the oxidative stress-induced neurodegeneration. Curr Alzheimer Res. 13:185–197. 2016. View Article : Google Scholar : PubMed/NCBI
9	Lu Z, Chouhan AK, Borycz JA, Lu Z, Rossano AJ, Brain KL, Zhou Y, Meinertzhagen IA and Macleod GT: High-probability neurotransmitter release sites represent an energy-efficient design. Curr Biol. 26:2562–2571. 2016. View Article : Google Scholar : PubMed/NCBI
10	Zhang Y and Bhavnani BR: Glutamate-induced apoptosis in neuronal cells is mediated via caspase-dependent and independent mechanisms involving calpain and caspase-3 proteases as well as apoptosis inducing factor (AIF) and this process is inhibited by equine estrogens. BMC Neurosci. 7:492006. View Article : Google Scholar : PubMed/NCBI
11	Lamigeon C, Bellier JP, Sacchettoni S, Rujano M and Jacquemont B: Enhanced neuronal protection from oxidative stress by coculture with glutamic acid decarboxylase-expressing astrocytes. J Neurochem. 77:598–606. 2001. View Article : Google Scholar : PubMed/NCBI
12	Ye ZC and Sontheimer H: Astrocytes protect neurons from neurotoxic injury by serum glutamate. Glia. 22:237–248. 1998. View Article : Google Scholar : PubMed/NCBI
13	Ha JS, Dho SH, Youm TH, Kwon KS and Park SS: Astrocytic phospholipase A2 contributes to neuronal glutamate toxicity. Brain Res 1590. 97–106. 2014. View Article : Google Scholar
14	Negishi T, Ishii Y, Kawamura S, Kuroda Y and Yoshikawa Y: Cryopreservation of brain tissue for primary culture. Exp Anim. 51:383–390. 2002. View Article : Google Scholar : PubMed/NCBI
15	Haseleu J, Anlauf E, Blaess S, Endl E and Derouiche A: Studying subcellular detail in fixed astrocytes: Dissociation of morphologically intact glial cells (DIMIGs). Front Cell Neurosci. 7:542013. View Article : Google Scholar : PubMed/NCBI
16	Zhou L, Li F, Xu HB, Luo CX, Wu HY, Zhu MM, Lu W, Ji X, Zhou QG and Zhu DY: Treatment of cerebral ischemia by disrupting ischemia-induced interaction of nNOS with PSD-95. Nat Med. 16:1439–1443. 2010. View Article : Google Scholar : PubMed/NCBI
17	Sun Y, März P, Otten U, Ge J and Rose-John S: The effect of gp130 stimulation on glutamate-induced excitotoxicity in primary hippocampal neurons. Biochem Biophys Res Commun. 295:532–539. 2002. View Article : Google Scholar : PubMed/NCBI
18	Bösel J, Gandor F, Harms C, Synowitz M, Harms U, Djoufack PC, Megow D, Dirnagl U, Hörtnagl H, Fink KB and Endres M: Neuroprotective effects of atorvastatin against glutamate-induced excitotoxicity in primary cortical neurones. J Neurochem. 92:1386–1398. 2005. View Article : Google Scholar : PubMed/NCBI
19	Jiang Q, Gu Z and Zhang G: Activation, involvement and nuclear translocation of c-Jun N-terminal protein kinase 1 and 2 in glutamate-induced apoptosis in cultured rat cortical neurons. Brain Res. 956:194–201. 2002. View Article : Google Scholar : PubMed/NCBI
20	Savaskan NE, Bräuer AU, Kühbacher M, Eyüpoglu IY, Kyriakopoulos A, Ninnemann O, Behne D and Nitsch R: Selenium deficiency increases susceptibility to glutamate-induced excitotoxicity. FASEB J. 17:112–114. 2003. View Article : Google Scholar : PubMed/NCBI
21	Maher P and Davis JB: The role of monoamine metabolism in oxidative glutamate toxicity. J Neurosci. 16:6394–6401. 1996. View Article : Google Scholar : PubMed/NCBI
22	Choi DW, Maulucci-Gedde M and Kriegstein AR: Glutamate neurotoxicity in cortical cell culture. J Neurosci. 7:357–368. 1987. View Article : Google Scholar : PubMed/NCBI
23	Amin N and Pearce B: Glutamate toxicity in neuron-enriched and neuron-astrocyte co-cultures: Effect of the glutamate uptake inhibitor L-trans-pyrrolidine-2,4-dicarboxylate. Neurochem Int. 30:271–276. 1997. View Article : Google Scholar : PubMed/NCBI
24	Hyndman AG: The effects of glutamate and kainate on cell proliferation in retinal cultures. Invest Ophthalmol Vis Sci. 25:558–563. 1984.PubMed/NCBI
25	Olney JW: Glutamate-induced neuronal necrosis in the infant mouse hypothalamus. An electron microscopic study. J Neuropathol Exp Neurol. 30:75–90. 1971. View Article : Google Scholar : PubMed/NCBI
26	Churn SB, Sombati S, Taft WC and DeLorenzo RJ: Excitotoxicity affects membrane potential and calmodulin kinase II activity in cultured rat cortical neurons. Stroke. 24:271–278. 1993. View Article : Google Scholar : PubMed/NCBI