6-hydroxydopamine (6-OHDA) Oxidative Stress Assay for Observing Dopaminergic Neuron Loss in Caenorhabditis elegans

Sarah-Lena Offenburger (Lead / Corresponding author), Anton Gartner

Research output: Contribution to journalArticle

Abstract

The nematode Caenorhabditis elegans is a powerful genetic model that can be used to investigate neuronal death. Research using C. elegans has been crucial to characterize cell death programmes that are conserved in mammals. Many neuronal signaling components, such as those mediating dopaminergic neurotransmission, are preserved as well. Dopaminergic neurons are progressively lost in Parkinson's disease and an important risk factor to develop this disease appears to be oxidative stress, the increased occurrence of highly reactive oxygen species. Oxidative stress-induced dopaminergic neurodegeneration is mimicked in animal models by treatment with 6-hydroxydopamine (6-OHDA), a dopamine analog, which is specifically taken up into dopaminergic neurons. After exposing C. elegans to 6-OHDA, the loss of fluorescently labeled dopaminergic neurons can be easily monitored. An organisms' sensitivity to oxidative stress is thought to be influenced by basal levels of intrinsic oxidative stress and the ability to counteract oxidative stress and oxidative stress-induced damage. The C. elegans '6-OHDA model' led to the discovery of novel genes that are required to protect dopaminergic neurons and it has helped to determine the effects of conserved cell death and cell engulfment pathways in dopaminergic neurodegeneration. Here, we describe a simple protocol that allows for the easy detection of dopaminergic neuron loss after 6-OHDA treatment in C. elegans.

LanguageEnglish
Article numbere3025
Number of pages12
JournalBio-Protocol
Volume8
Issue number18
DOIs
Publication statusPublished - 20 Sep 2018

Fingerprint

Oxidopamine
Dopaminergic Neurons
Caenorhabditis elegans
Oxidative Stress
Cell Death
Genetic Models
Genetic Association Studies
Synaptic Transmission
Parkinson Disease
Mammals
Reactive Oxygen Species
Dopamine
Animal Models
Research

Keywords

  • C. elegans
  • Caenorhabditis elegans
  • 6-OHDA
  • 6-hydroxydopamine
  • Oxidative stress assay
  • Intoxication
  • Dopaminergic neurodegeneration

Cite this

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title = "6-hydroxydopamine (6-OHDA) Oxidative Stress Assay for Observing Dopaminergic Neuron Loss in Caenorhabditis elegans",
abstract = "The nematode Caenorhabditis elegans is a powerful genetic model that can be used to investigate neuronal death. Research using C. elegans has been crucial to characterize cell death programmes that are conserved in mammals. Many neuronal signaling components, such as those mediating dopaminergic neurotransmission, are preserved as well. Dopaminergic neurons are progressively lost in Parkinson's disease and an important risk factor to develop this disease appears to be oxidative stress, the increased occurrence of highly reactive oxygen species. Oxidative stress-induced dopaminergic neurodegeneration is mimicked in animal models by treatment with 6-hydroxydopamine (6-OHDA), a dopamine analog, which is specifically taken up into dopaminergic neurons. After exposing C. elegans to 6-OHDA, the loss of fluorescently labeled dopaminergic neurons can be easily monitored. An organisms' sensitivity to oxidative stress is thought to be influenced by basal levels of intrinsic oxidative stress and the ability to counteract oxidative stress and oxidative stress-induced damage. The C. elegans '6-OHDA model' led to the discovery of novel genes that are required to protect dopaminergic neurons and it has helped to determine the effects of conserved cell death and cell engulfment pathways in dopaminergic neurodegeneration. Here, we describe a simple protocol that allows for the easy detection of dopaminergic neuron loss after 6-OHDA treatment in C. elegans.",
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note = "Wellcome trust grant to AG (0909444/Z/09/Z and a Parkinson's UK Grant (G0912) together with a Wellcome Trust Strategic award (097045/B/11/Z). We acknowledge the Dundee Imaging Facility which is supported by the Wellcome Trust Technology Platform award (097945/B/11/Z) and the MRC Next Generation Optical Microscopy award (MR/K015869/1). SLO was supported by a PhD from the Molecular and Cellular Biology programme funded by the Wellcome Trust and by ISSF funding from the Wellcome Trust. Some strains were provided by the CGC, which is funded by the NIH Office of Research Infrastructure Programs (P40 OD010440).",
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6-hydroxydopamine (6-OHDA) Oxidative Stress Assay for Observing Dopaminergic Neuron Loss in Caenorhabditis elegans. / Offenburger, Sarah-Lena (Lead / Corresponding author); Gartner, Anton.

In: Bio-Protocol, Vol. 8, No. 18, e3025, 20.09.2018.

Research output: Contribution to journalArticle

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