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Neurotoxicity Research

, Volume 35, Issue 3, pp 530–541 | Cite as

Kynurenine 3-Monooxygenase Activity in Human Primary Neurons and Effect on Cellular Bioenergetics Identifies New Neurotoxic Mechanisms

  • Gloria Castellano-Gonzalez
  • Kelly R. Jacobs
  • Emily Don
  • Nicholas J. Cole
  • Seray Adams
  • Chai K. Lim
  • David B. LovejoyEmail author
  • Gilles J. GuilleminEmail author
Original Article
  • 121 Downloads

Abstract

Upregulation of the kynurenine pathway (KP) of tryptophan metabolism is commonly observed in neurodegenerative disease. When activated, L-kynurenine (KYN) increases in the periphery and central nervous system where it is further metabolised to other neuroactive metabolites including 3-hydroxykynurenine (3-HK), kynurenic acid (KYNA) and quinolinic acid (QUIN). Particularly biologically relevant metabolites are 3-HK and QUIN, formed downstream of the enzyme kynurenine 3-monooxygenase (KMO) which plays a pivotal role in maintaining KP homeostasis. Indeed, excessive production of 3-HK and QUIN has been described in neurodegenerative disease including Alzheimer’s disease and Huntington’s disease. In this study, we characterise KMO activity in human primary neurons and identified new mechanisms by which KMO activation mediates neurotoxicity. We show that while transient activation of the KP promotes synthesis of the essential co-enzyme nicotinamide adenine dinucleotide (NAD+), allowing cells to meet short-term increased energy demands, chronic KMO activation induces production of reactive oxygen species (ROS), mitochondrial damage and decreases spare-respiratory capacity (SRC). We further found that these events generate a vicious-cycle, as mitochondrial dysfunction further shunts the KP towards the KMO branch of the KP to presumably enhance QUIN production. These mechanisms may be especially relevant in neurodegenerative disease as neurons are highly sensitive to oxidative stress and mitochondrial impairment.

Keywords

Kynurenine pathway Kynurenine 3-monooxygenase Oxidative stress Mitochondrial dysfunction 

Abbreviations

3-HK

3-Hydroxykynurenine

AA

Anthranilic acid

AD

Alzheimer’s disease

CNS

Central nervous system

DAPI

4,6-Diamindino-2-phenylindole

DCF

2-,7-Dichlorofluorescin

ETS

Electron transport system

FAD

Flavin adenine dinucleotide

FCCP

Carbonyl cyanide-4-(trifluoromethoxy)phenylhydrazone

H2O2

Hydrogen peroxide

HD

Huntington’s Disease

HPLC

High-performance liquid chromatography

IDO1

Indoleamine 2,3-dioxygenase 1

IDO2

Indoleamine 2,3-dioxygenase 2

INF-γ

Interferon gamma

KAT

Kynurenine aminotransferase

KMO

Kynurenine 3-monooxygenase

KP

Kynurenine pathway

KYN

Kynurenine

KYNA

Kynurenic acid

KYNU

Kynureninase

MR

Maximal respiration

NAD

Nicotinamide adenine dinucleotide

NADPH

Nicotinamide adenine dinucleotide phosphate

NMDA

N-methyl-D-aspartate

OCR

Oxygen consumption rate

PBS

Phosphate-buffered saline

QUIN

Quinolinic acid

R123

Rhodamine 123

ROS

Reactive oxygen species

ROX

Residual oxygen consumption/non-mitochondrial respiration

RT

Room temperature

SRC

Spare respiratory capacity

TDO

Tryptophan 2,3-dioxygenase

TRP

Tryptophan

UHPLC

Ultra-high performance liquid chromatography

α7nAChR

α-7 nicotinic acetyl choline receptor

ΔΨm

Mitochondrial membrane potential

Notes

Acknowledgements

This work was supported by the National Health and Medical Research Council (NHMRC), the Australian Research Council (ARC), Macquarie University, The Snow Foundation and the Ramaciotti Perpetual Foundation (Australia). Dr. Gloria Castellano-Gonzalez was a recipient of the Macquarie University international scholarship.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

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Copyright information

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

Authors and Affiliations

  • Gloria Castellano-Gonzalez
    • 1
  • Kelly R. Jacobs
    • 1
  • Emily Don
    • 2
  • Nicholas J. Cole
    • 2
  • Seray Adams
    • 1
  • Chai K. Lim
    • 1
  • David B. Lovejoy
    • 1
    Email author
  • Gilles J. Guillemin
    • 1
    Email author
  1. 1.Neuroinflammation Group, Centre for Motor Neuron Disease Research, Department of Biomedical Sciences, Faculty of Medicine and Health Sciences (FMHS)Macquarie UniversityNorth RydeAustralia
  2. 2.Centre for Motor Neuron Disease Research, Department of Biomedical Sciences, Faculty of Medicine and Health Sciences (FMHS)Macquarie UniversityNorth RydeAustralia

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