Proposal of an In Vivo Comet Assay Using Haemocytes of Drosophila melanogaster
Proposal of an In Vivo Comet Assay Using Haemocytes of Drosophila melanogaster
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DOI：
10.1002/em.20604
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发表时间：
2011-03-01
影响因子：
2.8
通讯作者：
Marcos, Ricardo
中科院分区：
文献类型：
--
作者：
Carmona, Erico R.;Guecheva, Temenouga N.;Marcos, Ricardo
这项研究提出了第一个应用程序的体内碱性彗星试验，使用血细胞的果蝇幼虫。这些细胞的作用类似于哺乳动物血液，可以很容易地获得，并代表了处理过的幼虫的总体暴露量。为了验证试验，我们评估了这些细胞对三种已知诱变剂的反应：甲磺酸乙酯（EMS）、重铬酸钾（PD）和γ辐射（γ辐射）。三龄果蝇幼虫暴露于不同浓度的EMS（1，2和4 mM）和PD（0.5，1和2.5 mM）和不同剂量的γ-照射（2，4和8 G γ）。随后，从幼虫中提取血淋巴，通过离心分离血细胞并用于彗星试验。血细胞表现出DNA损伤的显著剂量相关性增加，表明这些细胞对处理明显敏感。这些结果表明，建议在体内彗星试验，使用幼虫血细胞的D。可能是一个有用的体内试验的遗传毒性评估。Environ.摩尔变异体52：165-169，2011年。(C)2010 Wiley-Liss，Inc.
This study presents the first application of an in vivo alkaline comet assay using haemocytes of Drosophila melanogaster larvae. These cells, which play a role similar to that of mammalian blood, can be easily obtained and represent an overall exposure of the treated larvae. To validate the assay, we evaluated the response of these cells to three well-known mutagenic agents: ethyl methanesulfonate (EMS), potassium dichromate (PD), and gamma radiation (gamma-irradiation). Third-instar Drosophila larvae were exposed to different concentrations of EMS (1, 2, and 4 mM) and PD (0.5, 1, and 2.5 mM) and to different doses of gamma-irradiation (2, 4, and 8 G gamma). Subsequently, haemolymph was extracted from the larvae, and haemocytes were isolated by centrifugation and used in the comet assay. Haemocytes exhibited a significant dose-related increase in DNA damage, indicating that these cells are clearly sensitive to the treatments. These results suggest that the proposed in vivo comet test, using larvae haemocytes of D. melanogaster, may be a useful in vivo assay for genotoxicity assessment. Environ. Mol. Mutagen. 52: 165-169, 2011. (C) 2010 Wiley-Liss, Inc.