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FAM134B-Mediated ER-Phagy in Mg2+-Free Solution-Induced Mitochondrial Calcium Homeostasis and Cell Death in Epileptic Hippocampal Neurons

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Abstract

Mitochondrial-associated endoplasmic reticulum (ER) membranes (MAMs) regulate calcium (Ca2+) homeostasis via Ca2+ transport-related proteins such as inositol-1,4,5-triphosphate receptor (IP3R). FAM134B-mediated ER-phagy plays an important role in ER homeostasis. However, it remains unknown whether FAM134B-mediated ER-phagy affects mitochondrial Ca2+ homeostasis and cell death through MAMs. In this study, we demonstrated that colocalization degree of FAM134B with LC3 and the LC3-II/LC3-I ratio were elevated in the hippocampal neuronal culture (HNC) model of acquired epilepsy (AE), which indicate an increased level of autophagy. In this model, FAM134B overexpression enhanced ER-phagy, while FAM134B downregulation had the opposite effect. Additionally, FAM134B overexpression significantly reversed the increases in IP3R expression and mitochondrial Ca2+ concentration and the decrease in the ER Ca2+ concentration in this model. FAM134B overexpression also ameliorated the AE-induced ultrastructural damage in neuronal mitochondria, decrease in mitochondrial membrane potential (mMP), cytochrome c (CytC) release and caspase-3 activation, while FAM134B downregulation induced the opposite effects. Altogether, our data indicate that FAM134B-mediated ER-phagy can attenuate AE-induced neuronal apoptosis, possibly by modulating the IP3R in MAMs to alter Ca2+ exchange between ER and mitochondria and thus inhibit mitochondrial structural damage, a decrease in mMP, release of CytC and mitochondrial apoptosis.

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Funding

This study was supported by the National Natural Science Foundation of China [Nos. 81701272, 81971214]; Provincial Ministry Co-construction Project from Medical Scientific and Technological Research Program of Henan Province [No. SB201902011] and Training plan for young backbone teachers of Zhengzhou University [No. 2019ZDGGJS056].

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  1. Cui Wang and Yujuan Li have contributed equally to this work.

Authors and Affiliations

  1. Department of Clinical Laboratory, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, China

    Cui Wang, Jingyu Zhang, Nan Li, Xiaomei Hu, Wenjing Zhang & Liang Ming

  2. Department of Neurology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, China

    Yujuan Li, Yingjiao Li, Liyuan Du & Nanchang Xie

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  1. Cui Wang
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Contributions

NX and CW contributed to conception and design of the study; YL, YL, LD, JZ, NL, XH and WZ performed the experiments; NX and LM performed the statistical analysis; CW and YL wrote the manuscript. All authors contributed to manuscript revision read and approved the submitted version.

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Correspondence to Nanchang Xie or Liang Ming.

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Wang, C., Li, Y., Li, Y. et al. FAM134B-Mediated ER-Phagy in Mg2+-Free Solution-Induced Mitochondrial Calcium Homeostasis and Cell Death in Epileptic Hippocampal Neurons. Neurochem Res 46, 2485–2494 (2021). https://doi.org/10.1007/s11064-021-03389-9

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