Effect of glutamate on lysosomal membrane permeabilization in primary cultured cortical neurons

Yan,Min; Zhu,Wenbo; Zheng,Xiaoke; Li,Yuan; Tang,Lipeng; Lu,Bingzheng; Chen,Wenli; Qiu,Pengxin; Leng,Tiandong; Lin,Suizhen; Yan,Guangmei; Yin,Wei

doi:10.3892/mmr.2016.4819

March-2016 Volume 13 Issue 3

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March-2016 Volume 13 Issue 3

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Article Open Access

Effect of glutamate on lysosomal membrane permeabilization in primary cultured cortical neurons

Authors:
- Min Yan
- Wenbo Zhu
- Xiaoke Zheng
- Yuan Li
- Lipeng Tang
- Bingzheng Lu
- Wenli Chen
- Pengxin Qiu
- Tiandong Leng
- Suizhen Lin
- Guangmei Yan
- Wei Yin
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Affiliations: Department of Pharmacology, Zhongshan School of Medicine, Sun Yat‑sen University, Guangzhou, Guangdong 510080, P.R. China, Department of Pathology, The First Affiliated Hospital, Sun Yat‑sen University, Guangzhou, Guangdong 510089, P.R. China, Guangzhou Cellprotek Pharmaceutical Co., Ltd., Guangzhou, Guangdong 510663, P.R. China, Department of Biochemistry, Zhongshan School of Medicine, Sun Yat‑sen University, Guangzhou, Guangdong 510080, P.R. China

Copyright: © Yan et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
Pages: 2499-2505
|
Published online on: January 28, 2016

https://doi.org/10.3892/mmr.2016.4819
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Abstract

Glutamate is the principal neurotransmitter in the central nervous system. Glutamate-mediated excitotoxicity is the predominant cause of cerebral damage. Recent studies have shown that lysosomal membrane permeabilization (LMP) is involved in ischemia‑associated neuronal death in non‑human primates. This study was designed to investigate the effect of glutamate on lysosomal stability in primary cultured cortical neurons. Glutamate treatment for 30 min induced the permeabilization of lysosomal membranes as assessed by acridine orange redistribution and immunofluorescence of cathepsin B in the cytoplasm. Inhibition of glutamate excitotoxicity by the NMDA receptor antagonist MK‑801 and the calcium chelator ethylene glycol‑bis (2‑aminoethylether)‑N, N, N', N'‑tetraacetic acid, rescued lysosomes from permeabilization. The role of calpain and reactive oxygen species (ROS) in inducing LMP was also investigated. Ca2+ overload following glutamate treatment induced the activation of calpain and the production of ROS, which are two major contributors to neuronal death. It has been reported that lysosomal‑associated membrane protein 2 (LAMP2) and heat shock protein (HSP)70 are two calpain substrates that promote LMP in cancer cells; however, it was found that calpains were activated by glutamate, but only LAMP2 was subsequently degraded. Furthermore, LMP was not alleviated by treatment with the calpain inhibitors calpeptin and SJA6017, which blocked the cleavage of the calpain substrate α‑fodrin. It was demonstrated that LMP was significantly alleviated by treatment with the antioxidant N‑Acetyl‑L‑cysteine, indicating that LMP involvement in early glutamate excitotoxicity may be mediated partly by ROS rather than calpain activation. Overall, these data shed light on the role of ROS-mediated LMP in early glutamate excitotoxicity.

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