The application of the comet assay to assess the genotoxicity of environmental pollutants in the nematode Caenorhabditis elegans.
The application of the comet assay to assess the genotoxicity of environmental pollutants in the nematode Caenorhabditis elegans.
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DOI：
10.1016/j.etap.2016.06.020
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发表时间：
2016-07
影响因子：
4.3
通讯作者：
Arlt VM
中科院分区：
文献类型：
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作者：
Imanikia S;Galea F;Nagy E;Phillips DH;Stürzenbaum SR;Arlt VM
本研究建立了从C. elegans成立了。研究了苯并[a]芘（BaP）的遗传毒性。使用彗星试验评估线虫。BaP的彗星形成具有剂量依赖性。NER和BER相关的单链断裂都有助于BaP形成彗星。本研究旨在建立秀丽隐杆线虫（Caelophabditis elegans，C. elegans）的单细胞电泳试验（彗星试验）的碱性版本来评估环境污染物苯并[a]芘（BaP）的遗传毒性。在C. elegans（野生型[WT]; N2，布里斯托）暴露48 h后（0-40 μM）。BaP诱导彗星的浓度依赖性高达20 μM; 20 μM BaP时彗星尾DNA %为30%，对照组为10%。同样，BaP诱导的DNA损伤在C. DNA修复缺陷的线虫突变株。在xpa-1和apn-1突变体中，BaP诱导的彗星形成减少到WT背景水平，表明在彗星试验中检测到的BaP形成的损伤在核苷酸和碱基切除修复缺陷的细胞中分别不被识别。总之，我们的研究提供了一个协议，评估环境污染物的DNA损伤在整个线虫使用彗星试验。
A protocol for the isolation of cells from C. elegans was established. The genotoxicity of benzo[a]pyrene (BaP) in C. elegans was assessed using the comet assay. Comet formation by BaP was dose-dependent. Both NER- and BER-related single-strand breaks contribute to comet formation by BaP. This study aimed to establish a protocol for cell dissociation from the nematode Caenorhabditis elegans (C. elegans) to assess the genotoxicity of the environmental pollutant benzo[a]pyrene (BaP) using the alkaline version of the single cell electrophoresis assay (comet assay). BaP genotoxicity was assessed in C. elegans (wild-type [WT]; N2, Bristol) after 48 h exposure (0–40 μM). Induction of comets by BaP was concentration-dependent up to 20 μM; comet% tail DNA was ∼30% at 20 μM BaP and ∼10% in controls. Similarly, BaP-induced DNA damage was evaluated in C. elegans mutant strains deficient in DNA repair. In xpa-1 and apn-1 mutants BaP-induced comet formation was diminished to WT background levels suggesting that the damage formed by BaP that is detected in the comet assay is not recognised in cells deficient in nucleotide and base excision repair, respectively. In summary, our study provides a protocol to evaluate DNA damage of environmental pollutants in whole nematodes using the comet assay.