Analysis of accumulation of DNA double-strand breaks in mouse tissues by pulsed-field gel electrophoresis

IF 3.4 3区医学 Q2 PHARMACOLOGY & PHARMACY Toxicology and applied pharmacology Pub Date : 2025-02-20 DOI:10.1016/j.taap.2025.117278

Takeshi Terabayashi , Takako Sasaki , Toshimasa Ishizaki , Tadashi Tomo , Yoshihiro Nishida , Katsuhiro Hanada

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Abstract

Double-strand DNA breaks (DSBs) are among the most cytotoxic DNA lesions, which can lead to chromosomal instability and eventually cell death. The substances that can potentially induce DSB formation are thus regarded as genotoxic. To date, many genotoxicity tests for detecting DNA breaks have been designed. However, there are limited options available for measuring the accumulation of DSBs in vivo. In this study, we aimed to evaluate a method of detecting the DSBs formed by the direct action of genotoxic substances using pulsed-field gel electrophoresis (PFGE). This approach has the advantage of making it easier to distinguish between DSBs and single-strand DNA breaks (SSBs) induced by the direct action of genotoxic substances. To confirm the detection of DSBs using PFGE, we investigated their accumulation after treatment with cis-diamminedichloroplatinum(II) (cisplatin) or γ-rays in mouse organs. The results revealed the successful detection of cisplatin-induced DSB formation in mouse kidney and thymus and γ-ray-induced DSB formation in all organs. We also discuss the advantages of PFGE-based detection of DSBs in vivo.

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用脉冲场凝胶电泳分析小鼠组织中DNA双链断裂的积累

双链DNA断裂（DSBs）是最具细胞毒性的DNA病变之一，可导致染色体不稳定并最终导致细胞死亡。因此，可能诱发DSB形成的物质被认为是遗传毒性的。迄今为止，已经设计了许多用于检测DNA断裂的遗传毒性测试。然而，用于测量dsb在体内积累的方法有限。本研究旨在探讨一种利用脉冲场凝胶电泳（PFGE）检测遗传毒性物质直接作用形成的dsb的方法。这种方法的优点是更容易区分dsb和由遗传毒性物质直接作用引起的单链DNA断裂（SSBs）。为了证实PFGE对DSBs的检测，我们研究了顺式二胺氯铂（II）（顺铂）或γ射线处理后DSBs在小鼠器官中的积累情况。结果显示，在小鼠肾脏和胸腺中成功检测到顺铂诱导的DSB形成，在所有器官中成功检测到γ射线诱导的DSB形成。我们还讨论了基于pfge的dsb体内检测的优势。

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来源期刊

Toxicology and applied pharmacology 医学-毒理学

CiteScore

6.80

自引率

2.60%

发文量

309

审稿时长

32 days

期刊介绍： Toxicology and Applied Pharmacology publishes original scientific research of relevance to animals or humans pertaining to the action of chemicals, drugs, or chemically-defined natural products. Regular articles address mechanistic approaches to physiological, pharmacologic, biochemical, cellular, or molecular understanding of toxicologic/pathologic lesions and to methods used to describe these responses. Safety Science articles address outstanding state-of-the-art preclinical and human translational characterization of drug and chemical safety employing cutting-edge science. Highly significant Regulatory Safety Science articles will also be considered in this category. Papers concerned with alternatives to the use of experimental animals are encouraged. Short articles report on high impact studies of broad interest to readers of TAAP that would benefit from rapid publication. These articles should contain no more than a combined total of four figures and tables. Authors should include in their cover letter the justification for consideration of their manuscript as a short article.