The long amplicon quantitative PCR for DNA damage assay as a sensitive method of assessing DNA damage in the environmental model, Atlantic killifish (Fundulus heteroditus)
The long amplicon quantitative PCR for DNA damage assay as a sensitive method of assessing DNA damage in the environmental model, Atlantic killifish (Fundulus heteroditus)
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DOI：
10.1016/j.cbpc.2008.07.007
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发表时间：
2009-03-01
影响因子：
3.9
通讯作者：
Di Giulio, Richard T.
中科院分区：
文献类型：
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作者：
Jung, Dawoon;Cho, Youngeun;Di Giulio, Richard T.
DNA损伤是多种污染物的重要毒性机制，因此在生态毒理学研究中常被用作污染物效应的指标。在这里，我们采用了一种基于pcr的核和线粒体DNA损伤检测方法，用于一种重要的环境模型——大西洋鳉鱼（Fundulus heteroclitus）。我们将这种检测称为长扩增子定量PCR （LA-QPCR）检测。为了在鳉鱼身上验证这一方法，研究人员在给鱼注射10mg /kg苯并[a]芘的情况下，测量了鱼的肝脏、大脑和肌肉的DNA损伤。这种暴露导致0.4-0.8个病变/10 kb。我们还测量了居住在超级基金站点的鳉鱼肝脏和肌肉组织的DNA损伤，证实了这种方法在生物监测中的实用性。在这两种情况下，核DNA （nDNA）和线粒体DNA （mtDNA）的损伤水平相当。由于对于许多与环境相关的物种来说，广泛的nDNA序列数据并不容易获得，但线粒体基因组通常是完全测序的，因此这种测定方法可以适用于几乎任何没有发育的物种的mtDNA损伤。因此，我们认为这种分析将是评估生态毒理学研究中DNA损伤的有价值的工具。(C) 2008爱思唯尔公司版权所有。
DNA damage is an important mechanism of toxicity for a variety of pollutants, and therefore, is often used as an indicator of pollutant effects in ecotoxicological studies. Here, we adapted a PCR-based assay for nuclear and mitochondrial DNA damage for use in an important environmental model, the Atlantic killifish (Fundulus heteroclitus). We refer to this assay as the long amplicon quantitative PCR (LA-QPCR) assay. To validate this method in killifish, DNA damage was measured in liver, brain, and muscle of fish dosed with 10 mg/kg benzo [a]pyrene. This exposure caused 0.4-0.8 lesions/10 kb. We also measured DNA damage in liver and muscle tissues from killifish inhabiting a Superfund site, confirming the utility of this method for biomonitoring. In both cases, damage levels were comparable in nuclear DNA (nDNA) and mitochondrial DNA (mtDNA). Since extensive nDNA sequence data are not readily available for many environmentally relevant species, but mitochondrial genomes are frequently fully sequenced, this assay can be adapted to examine mtDNA damage in virtually any species with little development. Therefore, we argue that this assay will be a valuable tool in assessing DNA damage in ecotoxicological studies. (C) 2008 Elsevier Inc. All rights reserved.