Sykora Peter, Chiari Ylenia, Heaton Andrew, Moreno Nickolas, Glaberman Scott, Sobol Robert W
Department of Oncologic Sciences, Mitchell Cancer Institute, University of South Alabama, Mobile, Alabama, 36604.
Department of Biology, University of South Alabama, Mobile, Alabama, 36688.
Environ Mol Mutagen. 2018 May;59(4):322-333. doi: 10.1002/em.22183. Epub 2018 Mar 14.
DNA damage has been linked to genomic instability and the progressive breakdown of cellular and organismal homeostasis, leading to the onset of disease and reduced longevity. Insults to DNA from endogenous sources include base deamination, base hydrolysis, base alkylation, and metabolism-induced oxidative damage that can lead to single-strand and double-strand DNA breaks. Alternatively, exposure to environmental pollutants, radiation or ultra-violet light, can also contribute to exogenously derived DNA damage. We previously validated a novel, high through-put approach to measure levels of DNA damage in cultured mammalian cells. This new CometChip Platform builds on the classical single cell gel electrophoresis or comet methodology used extensively in environmental toxicology and molecular biology. We asked whether the CometChip Platform could be used to measure DNA damage in samples derived from environmental field studies. To this end, we determined that nucleated erythrocytes from multiple species of turtle could be successfully evaluated in the CometChip Platform to quantify levels of DNA damage. In total, we compared levels of DNA damage in 40 animals from two species: the box turtle (Terrapene carolina) and the red-eared slider (Trachemys scripta elegans). Endogenous levels of DNA damage were identical between the two species, yet we did discover some sex-linked differences and changes in DNA damage accumulation. Based on these results, we confirm that the CometChip Platform allows for the measurement of DNA damage in a large number of samples quickly and accurately, and is particularly adaptable to environmental studies using field-collected samples. Environ. Mol. Mutagen. 59:322-333, 2018. © 2018 Wiley Periodicals, Inc.
DNA损伤与基因组不稳定以及细胞和机体稳态的逐渐破坏有关，会导致疾病的发生和寿命缩短。内源性因素对DNA的损害包括碱基脱氨基、碱基水解、碱基烷基化以及代谢诱导的氧化损伤，这些都可能导致单链和双链DNA断裂。另外，接触环境污染物、辐射或紫外线也会造成外源性DNA损伤。我们之前验证了一种新的高通量方法来测量培养的哺乳动物细胞中的DNA损伤水平。这种新的彗星芯片平台是基于在环境毒理学和分子生物学中广泛使用的经典单细胞凝胶电泳或彗星试验方法构建的。我们想知道彗星芯片平台是否可用于测量来自环境现场研究的样本中的DNA损伤。为此，我们确定可以在彗星芯片平台上成功评估多种龟类的有核红细胞，以量化DNA损伤水平。我们总共比较了两种龟类（箱龟和红耳龟）的40只动物的DNA损伤水平。这两个物种的内源性DNA损伤水平相同，但我们确实发现了一些与性别相关的差异以及DNA损伤积累的变化情况。基于这些结果，我们证实彗星芯片平台能够快速、准确地测量大量样本中的DNA损伤，并且特别适用于使用现场采集样本的环境研究。《环境与分子诱变》，2018年，第59卷，第322 - 333页。© 2018威利期刊公司
