Integrated multi-omics analysis reveals the underlying molecular mechanism for developmental neurotoxicity of perfluorooctanesulfonic acid in zebrafish

Limited studies on multi-omics have been conducted to comprehensively investigate the molecular mechanism underlying the developmental neurotoxicity of perfluorooctanesulfonic acid (PFOS). In this study, the locomotor behavior of zebrafish larvae was assessed under the exposure to 0.1–20 µM PFOS based on its reported neurobehavioral effect. After the number of zebrafish larvae was optimized for proteomics and metabolomics studies, three kinds of omics (i.e., transcriptomics, proteomics, and metabolomics) were carried out with zebrafish larvae exposed to 0.1, 1, 5, and 10 µM PFOS. More importantly, a data-driven integration of multi-omics was performed to elucidate the toxicity mechanism involved in developmental neurotoxicity. In a concentration-dependent manner, exposure to PFOS provoked hyperactivity and hypoactivity under light and dark conditions, respectively. Individual omics revealed that PFOS exposure caused perturbations in the pathways of neurological function, oxidative stress, and energy metabolism. Integrated omics implied that there were decisive pathways for axonal deformation, neuroinflammatory stimulation, and dysregulation of calcium ion signaling, which are more clearly specified for neurotoxicity. Overall, our findings broaden the molecular understanding of the developmental neurotoxicity of PFOS, for which multi-omics and integrated omics analyses are efficient for discovering the significant molecular pathways related to developmental neurotoxicity in zebrafish. Overall design: Comparative gene expression profiling analysis of RNA-seq data for zebrafish (Danio_rerio)

目前针对全氟辛烷磺酸（perfluorooctanesulfonic acid，PFOS）发育性神经毒性的分子机制开展的多组学综合研究较为有限。本研究基于已报道的全氟辛烷磺酸神经行为毒性效应，采用0.1~20 μM浓度梯度的全氟辛烷磺酸暴露斑马鱼幼鱼，评估其运动行为变化。在优化了蛋白质组学与代谢组学研究所需的斑马鱼幼鱼数量后，本研究对暴露于0.1、1、5、10 μM全氟辛烷磺酸的斑马鱼幼鱼开展了转录组学、蛋白质组学及代谢组学三类组学分析。更为关键的是，本研究通过数据驱动的多组学整合分析，阐明了全氟辛烷磺酸发育性神经毒性相关的毒性机制。

暴露于全氟辛烷磺酸可呈浓度依赖性地分别在光照与黑暗条件下引发斑马鱼幼鱼的活动亢进与活动减退。单一组学分析结果显示，全氟辛烷磺酸暴露会干扰神经系统功能、氧化应激及能量代谢相关通路。整合组学分析则揭示了与神经毒性高度相关的关键通路，包括轴突变形、神经炎症刺激以及钙离子信号通路失调，这些通路与神经毒性的对应关系更为明确。

综上，本研究结果加深了人们对全氟辛烷磺酸发育性神经毒性分子机制的认知，同时证实多组学与整合组学分析可有效挖掘斑马鱼中与发育性神经毒性相关的关键分子通路。

实验整体设计：针对斑马鱼（Danio_rerio）的RNA测序（RNA-seq）数据开展比较基因表达谱分析。