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用于观察[具体部位]中多巴胺能神经元损失的6-羟基多巴胺(6-OHDA)氧化应激测定

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

作者信息

Offenburger Sarah-Lena, Gartner Anton

机构信息

Centre for Gene Regulation and Expression, School of Life Sciences, University of Dundee, Dundee, United Kingdom.

出版信息

Bio Protoc. 2018 Sep 20;8(18). doi: 10.21769/BioProtoc.3025.

Abstract

The nematode is a powerful genetic model that can be used to investigate neuronal death. Research using 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 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 '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 .

摘要

线虫是一种强大的遗传模型,可用于研究神经元死亡。利用该模型进行的研究对于表征在哺乳动物中保守的细胞死亡程序至关重要。许多神经元信号传导成分,如介导多巴胺能神经传递的成分,也得以保留。在帕金森病中,多巴胺能神经元会逐渐丧失,而氧化应激似乎是引发这种疾病的一个重要风险因素,氧化应激即高活性氧物种的发生率增加。在动物模型中,通过用多巴胺类似物6-羟基多巴胺(6-OHDA)处理可模拟氧化应激诱导的多巴胺能神经变性,6-OHDA会特异性地被多巴胺能神经元摄取。在用6-OHDA处理后,可轻松监测荧光标记的多巴胺能神经元的丧失情况。生物体对氧化应激的敏感性被认为受内在氧化应激的基础水平以及对抗氧化应激和氧化应激诱导损伤能力的影响。“6-OHDA模型”促使人们发现了保护多巴胺能神经元所需的新基因,并有助于确定保守的细胞死亡和细胞吞噬途径在多巴胺能神经变性中的作用。在此,我们描述了一种简单的方案,该方案可轻松检测经6-OHDA处理后的线虫中多巴胺能神经元的丧失情况。

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