α-Synuclein is involved in manganese-induced ER stress via PERK signal pathway in organotypic brain slice cultures.
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.
Department of Environmental Health, School of Public Health, China Medical University, Shenyang, Liaoning, 110001, People's Republic of China, xubin@mail.cmu.edu.cn.
Bin
B
Xu
Fei
F
Wang
Sheng-Wen
SW
Wu
Yu
Y
Deng
Wei
W
Liu
Shu
S
Feng
Tian-Yao
TY
Yang
Zhao-Fa
ZF
Xu
D000818:Animals; D000831:Animals, Newborn; D001921:Brain / Q000378:metabolism*; D059865:Endoplasmic Reticulum Stress / Q000502:physiology*; D008345:Manganese / Q000633:toxicity*; D009924:Organ Culture Techniques; D051381:Rats; D017208:Rats, Wistar; D015398:Signal Transduction / Q000502:physiology*; D051844:alpha-Synuclein / Q000502:physiology* / Q000633:toxicity; D019892:eIF-2 Kinase / Q000502:physiology*
D008345:Manganese; D051844:alpha-Synuclein; D019892:eIF-2 Kinase