Chaousis Stephanie, Leusch Frederic D L, Nouwens Amanda, Melvin Steven D, van de Merwe Jason P
Griffith School of Science and Environment and the Australian Rivers Institute, Griffith Univeristy, Building 51, Gold Coast Campus, QLD 4222, Australia.
School of Chemistry and Molecular Biology, The University of Queensland, Building 76, QLD 4067, Australia.
J Proteomics. 2023 Aug 15;285:104942. doi: 10.1016/j.jprot.2023.104942. Epub 2023 Jun 5.
Understanding the impacts of chemical exposure in marine wildlife is challenging, due to practical and ethical constraints that preclude traditional toxicology research on these animals. This study addressed some of these limitations by presenting an ethical and high throughput cell-based approach to elucidate molecular-level effects of contaminants on sea turtles. The experimental design addressed basic questions of cell-based toxicology, including chemical dose and exposure time. Primary green turtle skin cells were exposed to polychlorinated biphenyl (PCB) 153 and perfluorononanoic acid (PFNA) for 24 and 48 h, at three sub-lethal, environmentally relevant concentrations (1, 10 and 100 μg/L). Sequential window acquisition of all theoretical mass spectra (SWATH-MS) identified over 1000 differentially abundant proteins within the 1% false discovery rate (FDR) threshold. The 24 h exposure resulted in a greater number of differentially abundant proteins, compared to 48 h exposure, for both contaminants. However, there were no statistically significant dose-response relationships for the number of differentially synthesised proteins, nor differences in the proportion of increased vs decreased proteins between or within exposure times. Known in vivo markers of contaminant exposure, superoxide dismutase and glutathione S-transferase, were differentially abundant following exposure to PCB153 and PFNA. SIGNIFICANCE: Cell-based (in vitro) proteomics provides an ethical and high throughput approach to understanding the impacts of chemical contamination on sea turtles. Through investigating effects of chemical dose and exposure duration on unique protein abundance in vitro, this study provides an optimised framework for conducting cell-based studies in wildlife proteomics, and highlights that proteins detected in vitro could act as biomarkers of chemical exposure and effect in vivo.
由于实际和伦理限制阻碍了对这些动物进行传统毒理学研究，了解化学物质暴露对海洋野生动物的影响具有挑战性。本研究通过提出一种符合伦理且高通量的基于细胞的方法来阐明污染物对海龟的分子水平影响，解决了其中一些限制。实验设计解决了基于细胞的毒理学的基本问题，包括化学剂量和暴露时间。将绿海龟原代皮肤细胞暴露于多氯联苯（PCB）153和全氟壬酸（PFNA）中24小时和48小时，浓度为三个亚致死且与环境相关的浓度（1、10和100μg/L）。所有理论质谱的序列窗口采集（SWATH-MS）在1%错误发现率（FDR）阈值内鉴定出1000多种差异丰富的蛋白质。与48小时暴露相比，两种污染物在24小时暴露时产生的差异丰富蛋白质数量更多。然而，差异合成蛋白质的数量没有统计学上显著的剂量反应关系，暴露时间之间或之内增加与减少蛋白质的比例也没有差异。已知的污染物暴露体内标志物超氧化物歧化酶和谷胱甘肽S-转移酶在暴露于PCB153和PFNA后差异丰富。意义：基于细胞（体外）的蛋白质组学为理解化学污染对海龟的影响提供了一种符合伦理且高通量的方法。通过研究化学剂量和暴露持续时间对体外独特蛋白质丰度的影响，本研究为在野生动物蛋白质组学中进行基于细胞的研究提供了一个优化框架，并强调体外检测到的蛋白质可作为体内化学暴露和效应的生物标志物。
