Complementary Square Wave Voltammetry and Tandem Mass Spectrometry Analysis to Identify and Detect Compensatory Genomic Changes in Nematodes due to Nickel (II) Exposure
Complementary Square Wave Voltammetry and Tandem Mass Spectrometry Analysis to Identify and Detect Compensatory Genomic Changes in Nematodes due to Nickel (II) Exposure
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DOI：
10.1016/j.snr.2021.100070
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发表时间：
2021-12
影响因子：
5.9
通讯作者：
Elizabeth R. LaFave;Ryne Turner;Nicholas J. Schaaf;Thekra Hindi;David Rudel;Eli G. Hvastkovs
中科院分区：
文献类型：
--
作者：
Elizabeth R. LaFave;Ryne Turner;Nicholas J. Schaaf;Thekra Hindi;David Rudel;Eli G. Hvastkovs
镍是一种有毒的重金属，可能会导致负面的健康结果，包括接触后的癌症[1]。方波伏安法被用来检测DNA直接提取的镍暴露的线虫，起源于高或低含镍的环境，以评估进化遗传学在镍毒性过程中的作用。提取的DNA固定在热解石墨（PG）电极上，然后在Ru（bpy）3+存在下电化学氧化，在1.05 V vs. SCE下产生电催化氧化电流。BothC.雅丽根海峡使用太平洋线虫，每种线虫具有火山或世界性土壤来源菌株。DNA氧化峰电流（Ip）在暴露于50 μg/L Ni ~（2+）时增加，但与来自世界性土壤的线虫相比，来自火山土壤的线虫表现出显著（40-50%）低的Ipupon Ni ~（2+）暴露。进一步的SWV分析在来自一系列C. elegansN 2xCB 4856重组近交高级互交系群体（RIAL）。在RIAL菌株中，Ipmagnitude的连续性被视为Ni 2+暴露的函数，表明Ni耐受性是复杂的，并且受多个位点的影响。大部分的DNA从镍暴露的个体RIAL菌株培养物产生的氧化电流增加相比，DNA从类似的镍未暴露的文化相同的RIAL菌株。酸水解线虫DNA的液相色谱串联质谱（LC-MS/MS）分析提供了电化学研究结果的验证。鸟嘌呤和氧化损伤的鸟嘌呤含量通过适当的tem/zmass转换进行监测。鸟嘌呤含量在所有镍暴露RIAL DNA是较低的，而氧化损伤的鸟嘌呤升高相比，未暴露的线虫，但一个分析RIAL。相结合，互补的电化学和MS/MS数据提供的证据表明，进化遗传学导致遗传保护环境毒物，这表明可能有多个基因参与保护的有机体从镍暴露。
Nickel is a toxic heavy metal that may cause negative health outcomes including cancer upon exposure [1]. Square wave voltammetry was used to assay DNA directly extracted from nickel-exposed nematodes that had originated from either high or low Ni-containing environments in order to assess the role of evolutionary genetics in the Ni toxicity process. Extracted DNA was immobilized on pyrolytic graphite (PG) electrodes following layer by layer (LbL) protocols and then electrochemically oxidized in the presence of Ru(bpy)3+to generate electrocatalytic oxidative currents at ∼+1.05 V vs. SCE. BothC. elegansandP. pacificusnematodes were utilized, each with a volcanic or cosmopolitan soil source strain. DNA oxidative peak currents (Ip) increased for all nematode strains upon exposure to 50 μg/L Ni2+, but those originating from volcanic soils exhibited significantly (40–50%) lower Ipupon Ni2+exposure compared to similarly exposed nematodes from cosmopolitan soils. Further SWV analysis was performed on DNA from a series ofC. elegansN2xCB4856 recombinant inbred advanced intercross line populations (RIALs). A continuum of Ipmagnitude was seen as a function of Ni2+exposure among the RIAL strains indicating Ni-tolerance is complex and affected by multiple loci. The majority of the DNA from Ni-exposed individual RIAL strain cultures produced an increase in oxidative current in comparison to DNA from analogous Ni-unexposed cultures of the same RIAL strains. Liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis on acid hydrolyzed nematode DNA provided validation of the electrochemical findings. Guanine and oxidatively damaged guanine content were monitored via appropriatem/zmass transitions. Guanine content in all Ni-exposed RIAL DNA was lower, while oxidatively damaged guanine was elevated compared to unexposed nematodes in all but one analyzed RIAL. Combined, the complementary electrochemical and MS/MS data provide evidence that evolutionary genetics leads to genetic protection from environmental toxicants, suggesting a possibility that multiple genes are involved in the protection of the organism from Ni exposure.