Kober Sascha Liane, Meyer-Alert Henriette, Grienitz Desirée, Hollert Henner, Frohme Marcus
Molecular Biotechnology and Functional Genomics, Technical University of Applied Sciences Wildau, Hochschulring 1, 15745, Wildau, Germany.
Institute for Environmental Research (Biology V), Department of Ecosystem Analysis, RWTH Aachen University, Worringerweg 1, 52056, Aachen, Germany.
Anal Bioanal Chem. 2015 Oct;407(25):7721-31. doi: 10.1007/s00216-015-8937-2. Epub 2015 Aug 9.
In the last few decades, MALDI-TOF MS has become a useful technique not only in proteomics, but also as a fast and specific tool for whole cell analysis through intact cell mass spectrometry (IC-MS). The present study evaluated IC-MS as a novel tool for the detection of distinct patterns that can be observed after exposure to a certain toxin or concentration by utilizing the eukaryotic fish cell line RTL-W1. Two different viability assays were performed to define the range for IC-MS investigations, each of which employing copper sulfate, acridine, and β-naphthoflavone (BNF) as model compounds for several classes of environmental toxins. The IC-MS of RTL-W1 cells revealed not only specific spectral patterns for the various toxins, but also that the concentration used had an effect on RTL-W1 profiles. After the exposure with copper sulfate and acridine, the spectra of RTL-W1 showed a significant increase of certain peaks in the higher mass range (m/z >7000), which is probably attributed to the apoptosis of RTL-W1. On the contrary, exposure to BNF showed a distinct change of ion abundances only in the lower mass range (m/z <7000). Furthermore, a set of mass peaks could be identified as a specific biomarker for a single toxin treatment, so IC-MS demonstrates a new method for the distinction of toxic effects in fish cells. Due to fast sample preparation and high throughput, IC-MS offers great potential for ecotoxicological studies to investigate cellular effects of different substances and complex environmental samples.
在过去几十年中,基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)不仅在蛋白质组学中成为一种有用的技术,而且作为一种通过完整细胞质谱分析(IC-MS)进行全细胞分析的快速且特异的工具。本研究评估了IC-MS作为一种新型工具,通过利用真核鱼类细胞系RTL-W1来检测在暴露于特定毒素或浓度后可观察到的不同模式。进行了两种不同的活力测定以确定IC-MS研究的范围,每种测定都使用硫酸铜、吖啶和β-萘黄酮(BNF)作为几类环境毒素的模型化合物。RTL-W1细胞的IC-MS不仅揭示了各种毒素的特定光谱模式,还表明所用浓度对RTL-W1谱有影响。在用硫酸铜和吖啶处理后,RTL-W1的光谱显示在较高质量范围(m/z >7000)中某些峰显著增加,这可能归因于RTL-W1的凋亡。相反,暴露于BNF仅在较低质量范围(m/z <7000)显示离子丰度有明显变化。此外,一组质量峰可被鉴定为单一毒素处理的特定生物标志物,因此IC-MS证明了一种区分鱼类细胞中毒性效应的新方法。由于样品制备快速且通量高,IC-MS在生态毒理学研究中具有巨大潜力,可用于研究不同物质和复杂环境样品的细胞效应。