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BANF1 promotes glutamate-induced apoptosis of HT-22 hippocampal neurons

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Abstract

Background

Glutamate exposure was fatal to HT-22 neuronal cells that derived from mouse hippocampus. This is often used as a model for hippocampus neurodegeneration in vitro. The targets relevant to glutamate-induced neuronal toxicity is not fully understood. In this study, we aimed to identify crucial factors associated with glutamate-induced cytotoxicity in HT-22 cells.

Methods

HT-22 cells were treated with 7.5 mM glutamate for 24 h and isobaric tags for relative and absolute quantitation (iTRAQ) proteomic analysis conducted to identify the differentially expressed proteins. Differential proteins were subjected to Gene Ontology analyses. Upregulation of barrier to autointegration factor (BANF1/BANF1) protein was confirmed by RT-qPCR and western blotting. Cell viability was measured by CKK-8 and MTT assays. Cell apoptosis rates and intracellular reactive oxygen species (ROS) levels were detected using flow cytometry.

Results

A total of 5811 proteins were quantified by iTRAQ, 50 of which were recognized as significantly differential proteins (fold change ≥ 1.5 and P ≤ 0.05); 26 proteins were up-regulated and 24 were down-regulated after exposure to glutamate. GO enrichment analysis showed that the apoptotic signaling pathway was involved in cell death induced by glutamate. BANF1 expression level was markedly increased in HT-22 cells after glutamate treatment. Further, knockdown of BANF1 alleviated glutamate-mediated cell death with lower ROS levels.

Conclusions

In conclusion, we successfully filtered out differential proteins relevant to glutamate-mediated cytotoxicity. BANF1 upregulation promoted glutamate-induced apoptosis of HT-22 cells by enhancing ROS generation.

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Data availability

The data sets used and/or analyzed during the current study are available from the corresponding author on reasonable request privately.

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Funding

This study was funded by the Discipline Construction Project of Guangdong Medical University (4SG21229GDGFY01), the Science and Technology Program of Guangzhou (202103000051), and the Guangdong Provincial Natural Science Foundation General Project (2021A1515010780).

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Author notes

  1. Xinyu Yao and Xiaoyi Xu have contributed equally to this work.

Authors and Affiliations

  1. State Key Laboratory of Ophthalmology, Guangdong Provincial Key Laboratory of Ophthalmology and Visual Science, Zhongshan Ophthalmic Center, Sun Yat-Sen University, Guangzhou, 510060, China

    Xinyu Yao

  2. The First Dongguan Affiliated Hospital of Guangdong Medical University, Dongguan, 523710, Guangdong, China

    Shaodong Deng

  3. Scientific Research Platform, The Second Clinical Medical College, Guangdong Medical University, Dongguan, 523808, China

    Shaodong Deng

  4. The First Affiliated Hospital/School of Clinical Medicine of Guangdong Pharmaceutical University, Guangdong Pharmaceutical University, Guangzhou, 510080, China

    Zhaoshou Yang

  5. Bioland Laboratory, Guangzhou Regenerative Medicine and Health Guangdong Laboratory, Guangzhou, 510005, Guangdong, China

    Xiaoyi Xu

  6. Proteomics Research Center, Sun Yat-Sen Medical College of Sun Yat-Sen University, Guangzhou, 510080, China

    Kunhua Hu

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  1. Xinyu Yao
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Contributions

XY and KH performed the experiments and analyzed the data. XY and KH designed the study. XY and XY drafted the manuscript. SD and ZY supervised the study, reviewed and edited the manuscript. XY and XY contributed equally to this work and share first authorship.

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Correspondence to Zhaoshou Yang or Shaodong Deng.

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Yao, X., Xu, X., Hu, K. et al. BANF1 promotes glutamate-induced apoptosis of HT-22 hippocampal neurons. Mol Biol Rep 50, 9441–9452 (2023). https://doi.org/10.1007/s11033-023-08889-1

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