Grup de Mutagènesi, Departament de Genètica i de Microbiologia, Facultat de Biociències, Universitat Autònoma de Barcelona, Campus de Bellaterra, 08193 Cerdanyola del Vallès, Barcelona, Spain.
Mutat Res. 2011 Jan 10;718(1-2):33-7. doi: 10.1016/j.mrgentox.2010.10.008. Epub 2010 Nov 10.
In view of the scarcely available information on the in vivo mutagenic and co-mutagenic activity of nickel, the genotoxic potential of two nickel-compounds, nickel chloride (NiCl(2)) and nickel sulphate (NiSO(4)), was assessed in Drosophila melanogaster by measuring two different genetic endpoints. On the one hand, we used the wing-spot assay, which is based on the principle that the loss of heterozygosity of two suitable recessive markers, multiple wing hairs (mwh) and flare-3 (flr(3)), can lead to the formation of mutant clones in the imaginal disks of larval cells. On the other hand, the in vivo comet assay, which detects single- and double-strand DNA breaks, was also used with larval haemocytes. These cells offer several advantages: they are highly sensitive to genotoxic agents, the sampling and processing methodologies are quite simple and the level of basal DNA damage is relatively low. No significant increases in the frequencies of the three categories of mutant spots (i.e. small single spots, large single spots, and twin spots) were observed in the wing-spot assay; however, NiSO(4) induced significant dose-dependent increases in DNA damage in the comet assay. In addition, the combined treatments with gamma-radiation and NiCl(2) and NiSO(4) showed a slight but significant increase in the frequency of the three categories of mutant spots compared with the frequency induced by gamma-radiation alone, indicating that both nickel compounds have a synergistic interaction. These results support the assumption that both nickel compounds could act as co-mutagens interfering with DNA-repair processes and that the in vivo comet assay is a sensitive and effective method for detecting the DNA damage induced by NiSO(4) in haemocytes of D. melanogaster.
鉴于镍的体内致突变和协同致突变活性的信息很少,我们通过测量两个不同的遗传终点来评估两种镍化合物(氯化镍(NiCl(2))和硫酸镍(NiSO(4)))在黑腹果蝇中的遗传毒性潜力。一方面,我们使用翅斑测定法,该方法基于两个合适的隐性标记(多毛)和 flare-3(flr(3))的杂合性丧失可以导致幼虫细胞的成虫盘形成突变克隆的原理。另一方面,体内彗星测定法也用于检测幼虫血细胞中的单链和双链 DNA 断裂。这些细胞具有以下几个优点:它们对遗传毒性剂非常敏感,采样和处理方法相当简单,并且基础 DNA 损伤水平相对较低。在翅斑测定中,没有观察到三种突变斑(即小单斑、大单斑和双斑)频率的显著增加;然而,NiSO(4)在彗星测定中诱导了显著的剂量依赖性 DNA 损伤增加。此外,与单独γ辐射处理相比,γ辐射与 NiCl(2)和 NiSO(4)的联合处理显示三种突变斑频率略有但显著增加,表明两种镍化合物具有协同相互作用。这些结果支持以下假设:两种镍化合物可能作为协同致突变剂,干扰 DNA 修复过程,并且体内彗星测定法是检测 NiSO(4)在 D. melanogaster 血细胞中诱导的 DNA 损伤的一种敏感有效的方法。