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采用单细胞电感耦合等离子体质谱法和简易常压超声喷雾电离质谱法研究砷酸盐在细胞中的摄取及其对细胞脂质谱的影响。

Investigating the Uptake of Arsenate by Cells and its Effect on their Lipid Profile using Single Cell ICP-MS and Easy Ambient Sonic-Spray Ionization-MS.

机构信息

Environmental Chemical Processes Laboratory, Department of Chemistry , University of Crete , Voutes Campus, Heraklion 70013 , Greece.

Laboratory of Environmental Chemistry and of Biochemical Processes , Department of Agriculture, Hellenic Mediterranean University , Chania 73133 , Greece.

出版信息

Anal Chem. 2019 Aug 6;91(15):9590-9598. doi: 10.1021/acs.analchem.9b00917. Epub 2019 Jul 18.

Abstract

The complementary use of single cell atomic mass spectrometry (MS) and ambient molecular MS allowed for the in-depth study of arsenate uptake by cells and of the effect this toxic metalloid species has on their lipid profile. Compared to conventional inductively coupled plasma mass spectrometry (ICP-MS) analysis, in which case hundreds of thousands of cells are digested and then analyzed, it is demonstrated that single cell (SC) ICP-MS provides uptake data that are potentially of greater biological relevance. This includes the arsenic mass distribution within the cell population, which fits to a log-normal probability function, the most frequently contained arsenic mass within the cells (1.5-1.8 fg As per cell), and the mean arsenic uptake value (ranging from 2.7 to 4.1 fg As per cell for the three arsenate incubation concentrations, that is, 15, 22.5, and 30 μg As per mL) derived from the log-normal arsenic mass distribution within the cell population. The SC approach also allows for differentiating the arsenic present in and/or adsorbed on the cells, from the arsenic present in the extracellular solution, in a single analysis. In a similar fashion, ambient molecular MS in the form of desorption easy ambient sonic spray ionization (EASI) -MS was used to rapidly profile cell membrane lipids from cells spotted directly on a glass slide. EASI-MS analysis revealed that cells grown in the presence of increasing concentrations of arsenate exhibited changes in the degree of saturation of their membrane lipids, as was observed by the increasing intensity ratio of lipids with less unsaturated acyl chains to the same type of lipids with more unsaturated fatty acid chains. Thus, indicating "homeoviscous adaptation" of extraplastidial and thylakoid cell membranes, induced by the presence of arsenate.

摘要

单细胞原子质谱(MS)和环境分子 MS 的互补使用,使得深入研究细胞对砷酸盐的摄取以及这种有毒类金属物质对其脂质谱的影响成为可能。与传统的电感耦合等离子体质谱(ICP-MS)分析相比,在这种情况下,需要消化数十万计的细胞,然后进行分析,单细胞(SC)ICP-MS 提供的摄取数据具有更大的生物学相关性。这包括细胞群体中的砷质量分布,它符合对数正态概率函数,细胞中最常见的砷质量(每个细胞 1.5-1.8 fg As),以及细胞群体中对数正态砷质量分布所衍生的平均砷摄取值(对于三种砷酸盐孵育浓度,即 15、22.5 和 30 μg As/mL,范围从每个细胞 2.7 到 4.1 fg As)。SC 方法还允许在单次分析中区分存在于细胞内和/或吸附在细胞上的砷,以及存在于细胞外溶液中的砷。以同样的方式,以解吸简单环境超声喷雾电离(EASI)-MS 的形式的环境分子 MS 用于快速分析直接点在载玻片上的细胞的细胞膜脂质。EASI-MS 分析表明,在砷酸盐浓度不断增加的情况下生长的细胞,其膜脂质的饱和度发生了变化,这可以通过具有较少不饱和酰基链的脂质与具有更多不饱和脂肪酸链的相同类型的脂质的强度比增加来观察到。因此,这表明了由于砷酸盐的存在而诱导的质体外和类囊体细胞膜的“同型粘弹性适应”。

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