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Molecular Neurobiology

, Volume 55, Issue 8, pp 7025–7037 | Cite as

Disruption of De Novo Serine Synthesis in Müller Cells Induced Mitochondrial Dysfunction and Aggravated Oxidative Damage

  • Ting Zhang
  • Mark C. Gillies
  • Michele C. Madigan
  • Weiyong Shen
  • Jianhai Du
  • Ulrike Grünert
  • Fanfan Zhou
  • Michelle Yam
  • Ling Zhu
Article

Abstract

De novo serine synthesis plays important roles in normal mitochondrial function and cellular anti-oxidative capacity. It is reported to be mainly activated in glial cells of the central nervous system, but its role in retinal Müller glia remains unclear. In this study, we inhibited de novo serine synthesis using CBR-5884, a specific inhibitor of phosphoglycerate dehydrogenase (PHGDH, a rate limiting enzyme in de novo serine metabolism) in MIO-M1 cells (immortalized human Müller cells) and huPMCs (human primary Müller cells) under mild oxidative stress. Alamar blue and LDH (lactate dehydrogenase) assays showed significantly reduced metabolic activities and increased cellular damage of Müller cells, when exposed to CBR-5884 accompanied by mild oxidative stress; however, CBR-5884 alone had little effect. The increased cellular damage was partially reversed by supplementation with exogenous serine/glycine. HSP72 (an oxidative stress marker) and reactive oxygen species (ROS) levels were significantly increased; glutathione and NADPH/NADP+ levels were pronouncedly reduced under PHGDH inhibition accompanied by oxidative stress. JC-1 staining and Seahorse respiration experiments showed that inhibition of de novo serine synthesis in Müller cells can also increase mitochondrial stress and decrease mitochondrial ATP production. qPCR and Western blot demonstrated an increased expression of HSP60 (a key mitochondrial stress-related gene), and this was further validated in human retinal explants. Our study suggests that de novo serine synthesis is important for Müller cell survival, particularly when they are exposed to mild oxidative stress, possibly by maintaining mitochondrial function and generating glutathione and NADPH to counteract ROS.

Keywords

Müller cells De novo serine synthesis Mitochondrial dysfunction Oxidative stress Phosphoglycerate dehydrogenase Glutathione 

Notes

Acknowledgments

This study is supported by a grant from the Lowy Medical Research Institute. Professor Mark C. Gillies is a Sydney Medical School Fellow and is supported by a NHMRC Practitioner Fellowship. This paper formed the foundation of NHMRC project grant APP1145121.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

Supplementary material

12035_2017_840_MOESM1_ESM.pdf (212 kb)
ESM 1 (PDF 211 kb)

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2017

Authors and Affiliations

  1. 1.State Key Laboratory of Biotherapy and Cancer Center, Collaborative Innovation Center for Biotherapy, West China HospitalSichuan UniversityChengduPeople’s Republic of China
  2. 2.Save Sight InstituteThe University of SydneySydneyAustralia
  3. 3.School of Optometry and Vision SciencesUniversity of New South WalesSydneyAustralia
  4. 4.West Virginia University Health Sciences CenterMorgantownUSA
  5. 5.Faculty of PharmacyThe University of SydneySydneyAustralia

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