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Energy Metabolism and Mitochondrial Superoxide Anion Production in Pre-symptomatic Striatal Neurons Derived from Human-Induced Pluripotent Stem Cells Expressing Mutant Huntingtin

  • James Hamilton
  • Tatiana Brustovetsky
  • Akshayalakshmi Sridhar
  • Yanling Pan
  • Theodore R. Cummins
  • Jason S. Meyer
  • Nickolay BrustovetskyEmail author
Article
First Online:

Abstract

In the present study, we investigated whether mutant huntingtin (mHTT) impairs mitochondrial functions in human striatal neurons derived from induced pluripotent stem cells (iPSCs). Striatal neurons and astrocytes derived from iPSCs from unaffected individuals (Ctrl) and Huntington’s disease (HD) patients with HTT gene containing increased number of CAG repeats were used to assess the effect of mHTT on bioenergetics and mitochondrial superoxide anion production. The human neurons were thoroughly characterized and shown to express MAP2, DARPP32, GABA, synapsin, and PSD95. In human neurons and astrocytes expressing mHTT, the ratio of mHTT to wild-type huntingtin (HTT) was 1:1. The human neurons were excitable and could generate action potentials, confirming successful conversion of iPSCs into functional neurons. The neurons and astrocytes from Ctrl individuals and HD patients had similar levels of ADP and ATP and comparable respiratory and glycolytic activities. The mitochondrial mass, mitochondrial membrane potential, and superoxide anion production in human neurons appeared to be similar regardless of mHTT presence. The present results are in line with the results obtained in our previous studies with isolated brain mitochondria and cultured striatal neurons from YAC128 and R6/2 mice, in which we demonstrated that mutant huntingtin at early stages of HD pathology does not deteriorate mitochondrial functions. Overall, our results argue against bioenergetic deficits as a factor in HD pathogenesis and suggest that other detrimental processes might be more relevant to the development of HD pathology.

Keywords

Huntington’s disease Mitochondria Neurons Respiration Glycolysis Reactive oxygen species 

Abbreviations

mHTT

Mutant huntingtin

HTT

Wild-type huntingtin

iPSCs

Induced pluripotent stem cells

neurons

Human medium spiny neurons

HD

Huntington’s disease

ROS

Reactive oxygen species

BDNF

Brain-derived neurotrophic factor

GDNF

Glia-derived neurotrophic factor

DAPI

2-(4-Amidinophenyl)-1H-indole-6-carboxamidine

OCRs

Oxygen consumption rates

ECARs

Extracellular acidification rates

TMRM

Tetramethylrhodamine, methyl ester

TH

Tyrosine hydroxylase

AP

Action potentials

2,4-DNP

2,4-Dinitrophenol

FCCP

Carbonyl cyanide p-trifluoromethoxyphenylhydrazone

(hESCs)

Human embryonic stem cells

polyQ

Poly-glutamine

Ant

Antimycin A

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Notes

Acknowledgments

We are very thankful to Dr. George Daley (Harvard University, Cambridge, MA) and Dr. David Gamm (University of Wisconsin, Madison, WI) for providing human undifferentiated induced pluripotent stem cells.

Funding Information

This study was supported by National Institutes of Health grant R01 NS098772 and in part by a grant from Indiana Traumatic Spinal Cord & Brain Injury Research Fund to N.B.

Compliance with Ethical Standards

Conflict of Interests

The authors declare that they have no conflict of interests.

Supplementary material

12035_2019_1734_MOESM1_ESM.docx (12.4 mb)
ESM 1 (DOCX 12674 kb)

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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Pharmacology and Toxicology, School of MedicineIndiana UniversityIndianapolisUSA
  2. 2.Program in Medical Neuroscience, School of MedicineIndiana UniversityIndianapolisUSA
  3. 3.Paul and Carole Stark Neurosciences Research Institute, School of MedicineIndiana UniversityIndianapolisUSA
  4. 4.Department of Biology, School of ScienceIUPUIIndianapolisUSA
  5. 5.Indiana University School of MedicineIndianapolisUSA

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