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Journal of Molecular Neuroscience

, Volume 66, Issue 2, pp 229–237 | Cite as

Nrf2 Signaling in Sodium Azide-Treated Oligodendrocytes Restores Mitochondrial Functions

  • Annette Liessem-Schmitz
  • Nico Teske
  • Miriam Scheld
  • Stella Nyamoya
  • Adib Zendedel
  • Cordian Beyer
  • Tim Clarner
  • Athanassios Fragoulis
Article

Abstract

Mitochondrial dysfunctions mark a critical step in many central nervous system (CNS) pathologies, including multiple sclerosis (MS). Such dysfunctions lead to depolarization of mitochondrial membranes and imbalanced redox homeostasis. In this context, reactive oxygen species (ROS) are potentially deleterious but can also act as an important signaling step for cellular maintenance. The transcription factor nuclear factor (erythroid-derived 2)-like 2 (Nrf2), the key regulator in the cellular oxidative stress-response, induces a battery of genes involved in repair and regeneration. Here, we investigated the relevance of Nrf2 signaling for the prevention of cellular damage caused by dysfunctional mitochondria. We employed sodium azide (SA) as mitochondrial inhibitor on oligodendroglial OliNeu cells in vitro, and the cuprizone model with wild type and GFAP-Cre+::Keap1loxP/loxP mice to induce mitochondrial defects. The importance of Nrf2 for cellular functions and survival after SA treatment was elucidated by in vitro knockdown experiments with shRNA directed against Nrf2 and its inhibitor Keap1 as well as by methysticin treatment. Metabolic activity, cytotoxicity, and depolarization of the mitochondrial membrane were analyzed after SA treatment. The expression of Nrf2 target genes as well as endoplasmic reticulum stress response genes was additionally measured by real-time PCR (in vitro) and PCR gene arrays (in vivo). Treatment of OliNeu cells with SA resulted in significant depolarization of the mitochondrial membrane, decreased metabolic activity, and increased cytotoxicity. This was partly counteracted in Nrf2-hyperactivated cells and intensified in Nrf2-knockdown cells. Our studies demonstrate a key role of Nrf2 in maintaining cellular functions and survival in the context of mitochondrial dysfunction.

Keywords

Nrf2 Oligodendrocytes Mitochondrial dysfunction Complex IV Sodium azide Depolarization 

Notes

Acknowledgements

We thank Helga Helten, Petra Ibold, and Uta Zahn for their excellent technical assistance. We thank Sandra Amor for her helpful input.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

Supplementary material

12031_2018_1159_MOESM1_ESM.pdf (3.1 mb)
Fig. S1 The Nrf2/Keap1 axis and applied shRNA strategies for KD experiments. (left side) Under steady-state conditions, Nrf2 is scavenged by its intracellular inhibitor Keap1. This interaction leads directly to 26S proteasome-mediated degradation of Nrf2. In the presence of Nrf2 activating stimuli, Keap1 dissociates from Nrf2 and enables its nuclear translocation. There, Nrf2 binds to the anti-oxidant response elements (ARE) within the promoter region of its target genes and thereby induces or enhances their gene expression. (right side) In our experiments, we applied shRNA against Keap1 to decrease Keap1 protein content and thereby boost the Nrf2 activation in OliNeu cells even under steady-state conditions. The use of shRNA directed against Nrf2 was chosen to decrease Nrf2 activity in OliNeu cells. (PDF 3138 kb)
12031_2018_1159_MOESM2_ESM.xlsx (33 kb)
ESM 1 (XLSX 32 kb)

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Institute of NeuroanatomyUniklinik RWTH Aachen UniversityAachenGermany
  2. 2.Department of Anatomy IILudwig-Maximilians-University of MunichMunichGermany
  3. 3.Department of Anatomy and Cell Biology, Medical FacultyUniklinik RWTH Aachen UniversityAachenGermany

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