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Alpha-Synuclein is Involved in Manganese-Induced ER Stress via PERK Signal Pathway in Organotypic Brain Slice Cultures

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Abstract

Overexposure to manganese (Mn) has been known to induce neuronal damage involving endoplasmic reticulum (ER) stress. However, the exact mechanism of Mn-induced ER stress is unclear. Increasing evidence suggested that the overexpression of alpha-synuclein played a critical role in Mn-induced neurotoxicity. To explore whether the occurrence of ER stress was associated with alpha-synuclein overexpression, we made the rat brain slices model of silencing alpha-synuclein using short-interference RNA. After non-silencing alpha-synuclein slices were treated with Mn (0–400 μM) for 24 h, there was a dose-dependent increase in apoptotic rates of cells and levels of lactate dehydrogenase in the culture medium. Moreover, there was a dose-dependent increase in the protein expression of 78, 94-kDa glucose-regulated protein (GRP78/94), C/EBP homologous protein (CHOP), and caspase-12. Moreover, PKR-like ER kinase (PERK) phosphorylation, PERK-mediated phosphorylation of eIF2a, and ATF4 expression also increased. Inositol-requiring enzyme 1 (IRE1) activation and X-box-binding protein-1 (Xbp1) mRNA splicing increased. Activating transcription factor 6 p90 levels did not change. However, after silencing alpha-synuclein slices were treated with 400 μM Mn for 24 h, there was a significant decrease in the expression of GRP78/94, CHOP, and caspase-12 compared with 400 μM Mn-treated non-silencing alpha-synuclein slices. Furthermore, PERK phosphorylation, PERK-mediated phosphorylation of eIF2a, and ATF4 mRNA expression also decreased. However, IRE1 phosphorylation and Xbp1 mRNA splicing did not change. The findings revealed that Mn induced ER stress via activation of PERK and IRE1 signaling pathways and subsequent apoptosis in cultured slices. Moreover, alpha-synuclein protein was associated with Mn-induced activation of PERK signaling pathway.

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Acknowledgments

This work was supported by grants from the National Natural Science Foundation of China (grant no. 81102098).

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Environmental Health, School of Public Health, China Medical University, Shenyang, Liaoning, 110001, People’s Republic of China

    Bin Xu, Yu Deng, Wei Liu, Shu Feng, Tian-Yao Yang & Zhao-Fa Xu

  2. Department of Infection Management, Shengjing Hospital Affiliated to China Medical University, Shenyang, People’s Republic of China

    Fei Wang

  3. Department of Toxicology, School of Public Health, China Medical University, Shenyang, People’s Republic of China

    Sheng-Wen Wu

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  1. Bin Xu
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Correspondence to Bin Xu.

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Xu, B., Wang, F., Wu, SW. et al. Alpha-Synuclein is Involved in Manganese-Induced ER Stress via PERK Signal Pathway in Organotypic Brain Slice Cultures. Mol Neurobiol 49, 399–412 (2014). https://doi.org/10.1007/s12035-013-8527-2

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  • DOI: https://doi.org/10.1007/s12035-013-8527-2

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