Skip to main content
SpringerLink
Log in

Activating Transcription Factor 4-mediated Mitochondrial Unfolded Protein Response Alleviates Hippocampal Neuronal Damage in an In Vitro Model of Epileptiform Discharges

  • Original Paper
  • Published:
Neurochemical Research Aims and scope Submit manuscript

Abstract

The mitochondrial unfolded protein response (mtUPR) has been shown to restore protein homeostasis and cell function under stress, and recent studies have confirmed that the activating transcription factor 4 (ATF4) regulates mtUPR. However, the role of ATF4-mediated mtUPR in a hippocampal neuronal culture model of seizures remains unclear. Our results showed that the expression of mtUPR-related proteins (HSP60 and CLpP) increased in primary hippocampal neurons with seizures induced by a magnesium-free solution, suggesting mtUPR activation. Furthermore, ATF4 overexpression by lentiviral vector transfection enhanced the expression of HSP60 and CLpP, whereas ATF4 low expression by lentiviral vector transfection weakened the expression of HSP60 and CLpP. In addition, ATF4 overexpression increased neuronal viability and reduced seizure-induced apoptosis. ATF4 overexpression reduced reactive oxygen species (ROS) production and improved mitochondrial membrane potential damage during seizures. Moreover, ATF4 overexpression reduced the BCL2-associated X protein (Bax) expression and increased the expression of B-cell lymphoma 2 (BCL2). In contrast, ATF4 expression showed the opposite trend. In conclusion, our results showed that ATF4-mediated mtUPR may delay the cascade activation of apoptotic pathways by reducing ROS-mediated oxidative stress, thereby attenuating seizure-induced stress injury.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

Data Availability

All data generated by the current study are available upon reasonable request.

Code Availability

Not applicable.

References

  1. Misrani A, Tabassum S, Li Y (2021). Mitochondrial dysfunction and oxidative stress in Alzheimer’s Disease[J].Frontiers in Aging Neuroscience,13

  2. Shamima R (2018) Mitochondrial diseases and status epilepticus[J].Epilepsia,

  3. Lax NZ, Gorman GS, Turnbull DM (2017) Review: Central nervous system involvement in mitochondrial disease. Neuropathol Appl Neurobiol 43(2):102–118. https://doi.org/10.1111/nan.12333

    Article  CAS  PubMed  Google Scholar 

  4. Bindoff LA, Engelsen BA (2012) Mitochondrial diseases and epilepsy. Epilepsia 53(Suppl 4):92–97. https://doi.org/10.1111/j.1528-1167.2012.03618.x

    Article  CAS  PubMed  Google Scholar 

  5. Naresh NU, Haynes CM (2019) Signaling and regulation of the mitochondrial unfolded protein Response[J]. Cold Spring Harbor perspectives in biology

  6. Svagua T, Martini M, Martini M et al (2020) Mitochondrial unfolded protein response, mitophagy and other mitochondrial quality control mechanisms in heart disease and aged heart[J]. Croatian Med J 61(2):126–138

    Article  Google Scholar 

  7. Hu D, Liu Z, Qi X (2021) UPR(mt) activation protects against MPP(+)-induced toxicity in a cell culture model of Parkinson’s disease. Biochem Biophys Res Commun 569:17–22

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Melber A, Haynes CM (2018) UPR(mt) regulation and output: a stress response mediated by mitochondrial-nuclear communication. Cell Res 28(3):281–295

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Wortel IMN, van der Meer LT, Kilberg MS, van Leeuwen FN (2017) Surviving stress: modulation of ATF4-Mediated stress responses in normal and malignant cells. Trends Endocrinol Metab 28(11):794–806

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Quirós PM, Prado MA, Zamboni N, D’Amico D, Williams RW, Finley D, Gygi SP, Auwerx J Multi-omics analysis identifies ATF4 as a key regulator of the mitochondrial stress response in mammals.The Journal of cell biology2017, 216 (7),2027–2045

  11. Nargund AM, Fiorese CJ, Pellegrino MW, Deng P, Haynes CM (2015) Mitochondrial and nuclear accumulation of the transcription factor ATFS-1 promotes OXPHOS recovery during the UPR(mt). Mol Cell 58(1):123–133

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Jiang D, Cui H, Xie N, Banerjee S, Liu RM, Dai H, Thannickal VJ, Liu G (2020) ATF4 mediates mitochondrial unfolded protein response in alveolar epithelial cells. Am J Respir Cell Mol Biol 63(4):478–489

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Brewer GJ, Torricelli JR, Evege EK, Price PJ (1993) Optimized survival of hippocampal neurons in B27-supplemented Neurobasal, a new serum-free medium combination. J Neurosci Res 35(5):567–576

    Article  CAS  PubMed  Google Scholar 

  14. Sombati S, Delorenzo RJ (1995) Recurrent spontaneous seizure activity in hippocampal neuronal networks in culture. J Neurophysiol 73(4):1706–1711

    Article  CAS  PubMed  Google Scholar 

  15. Mackenzie-Gray Scott C, Parrish RR, Walsh D, Racca C, Cowell RM, Trevelyan AJ (2022) PV-specific loss of the transcriptional coactivator PGC-1α slows down the evolution of epileptic activity in an acute ictogenic model. J Neurophysiol. Jan 1;127(1):86–98. doi: 10.1152/jn.00295.2021. Epub 2021 Nov 17. PMID: 34788174; PMCID: PMC8721902

  16. Rho JM, Boison D (2022) The metabolic basis of epilepsy. Nat Rev Neurol 18(6):333–347. https://doi.org/10.1038/s41582-022-00651-8

    Article  CAS  PubMed  Google Scholar 

  17. Yang N, Guan QW, Chen FH et al (2020) Antioxidants targeting mitochondrial oxidative stress: promising Neuroprotectants for Epilepsy. Oxid Med Cell Longev. 2020:6687185. Published 2020 Nov 25 https://doi.org/10.1155/2020/6687185

    Article  PubMed  PubMed Central  Google Scholar 

  18. Wang X, Yu X, Li Y et al (2022) ATF5 attenuates apoptosis in hippocampal neurons with seizures evoked by Mg2+-Free medium via regulating mitochondrial unfolded protein response [published online ahead of print, 2022 Aug 8. Neurochem Res. https://doi.org/10.1007/s11064-022-03702-0

    Article  PubMed  PubMed Central  Google Scholar 

  19. Jin SM, Youle RJ (2013) The accumulation of misfolded proteins in the mitochondrial matrix is sensed by PINK1 to induce PARK2/Parkin-mediated mitophagy of polarized mitochondria. Autophagy 9(11):1750–1757

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Cappello F, Conway de Macario E, Marasà L, Zummo G, Macario AJ (2008) Hsp60 expression, new locations, functions and perspectives for cancer diagnosis and therapy. Cancer Biol Ther 7(6):801–809

    Article  CAS  PubMed  Google Scholar 

  21. Emre S (2019) Tianren, et al. Mitochondrial unfolded protein response: a stress response with implications for fertility and reproductive aging.[J]. Fertility and sterility

  22. Singh S, Singh TG, Rehni AK, Sharma V, Singh M, Kaur R (2021) Reviving mitochondrial bioenergetics: a relevant approach in epilepsy. Mitochondrion 58:213–226

    Article  CAS  PubMed  Google Scholar 

  23. Zsurka G, Kunz WS (2015) Mitochondrial dysfunction and seizures: the neuronal energy crisis. Lancet Neurol 14(9):956–966

    Article  CAS  PubMed  Google Scholar 

  24. Vovk T, Grabnar I, Martinc B (2014) Antioxidants as a preventive treatment for epileptic process: a review of the current Status[J]. Curr Neuropharmacol 12(6):–

  25. Sinha K, Das J, Pal PB et al (2013) Oxidative stress: the mitochondria-dependent and mitochondria-independent pathways of apoptosis[J]. Arch Toxicol 87(7):1157–1180

    Article  CAS  PubMed  Google Scholar 

  26. Valentin R, Grabow S, Davids MS (2018) The rise of apoptosis: targeting apoptosis in hematologic malignancies. Blood 132(12):1248–1264

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We acknowledge the support of the Translational Medicine Platform of Academy of Medical Sciences, Zhengzhou University.

Funding

This work was supported by National Natural Science Foundation of China (81971214) and the Outstanding Young Talent Cultivation Project of Henan Science and Technology Innovation Talents(YXKC2022037).

Author information

Author notes

  1. Xiaomeng Yu and Xiaoyi Wang contributed equally to this work.

Authors and Affiliations

  1. Department of Neurology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, China

    Xiaomeng Yu, Yinyin Xie, Tingting Peng, Yajun Lian, Cui Wang & Nanchang Xie

  2. Department of Clinical Laboratory, Key Clinical Laboratory of Henan Province, the First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, China

    Xiaoyi Wang, Cui Wang & Nanchang Xie

Authors
  1. Xiaomeng Yu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xiaoyi Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yinyin Xie
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Tingting Peng
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yajun Lian
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Cui Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Nanchang Xie
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

All authors contributed to the study conception and design.XMY,XYW, and YYX performed the experiments. XMY, XYW, and TTP analyzed the data. The first draft of the manuscript was written by XMY and XYW. CW and NCX contributed to the study conception and design, and revised the manuscript. All authors have read and approved the final manuscript.

Corresponding authors

Correspondence to Cui Wang or Nanchang Xie.

Ethics declarations

Competing Interests

All other authors declare that there is no potential conflict of interest.

Consent to Participate

Not applicable.

Consent for Publication

Not applicable.

Ethical Statement

All methods were performed carried out in accordance with relevant guidelines and regulations and were approved by the Animal Care and Use Commission of Zhengzhou University.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic Supplementary Material

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Yu, X., Wang, X., Xie, Y. et al. Activating Transcription Factor 4-mediated Mitochondrial Unfolded Protein Response Alleviates Hippocampal Neuronal Damage in an In Vitro Model of Epileptiform Discharges. Neurochem Res 48, 2253–2264 (2023). https://doi.org/10.1007/s11064-023-03910-2

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11064-023-03910-2

Keywords

Search

Navigation