BACKGROUND Per- and polyfluoroalkyl Substances (PFAS) are synthetic chemicals widely detected in humans and the environment. Exposure to perfluorooctanesulfonic acid (PFOS) or perfluorohexanesulfonic acid (PFHxS) was previously shown to cause dark-phase hyperactivity in larval zebrafish. OBJECTIVES The objective of this study was to elucidate the mechanism by which PFOS or PFHxS exposure caused hyperactivity in larval zebrafish. METHODS Swimming behavior was assessed in 5-d postfertilization (dpf) larvae following developmental (1-4 dpf) or acute (5 dpf) exposure to 0.43-7.86μM PFOS, 7.87-120μM PFHxS, or 0.4% dimethyl sulfoxide (DMSO). After developmental exposure and chemical washout at 4 dpf, behavior was also assessed at 5-8 dpf. RNA sequencing was used to identify differences in global gene expression to perform transcriptomic benchmark concentration-response (BMCT) modeling, and predict upstream regulators in PFOS- or PFHxS-exposed larvae. CRISPR/Cas9-based gene editing was used to knockdown peroxisome proliferator-activated receptors (ppars) pparaa/ab, pparda/db, or pparg at day 0. Knockdown crispants were exposed to 7.86μM PFOS or 0.4% DMSO from 1-4 dpf and behavior was assessed at 5 dpf. Coexposure with the ppard antagonist GSK3787 and PFOS was also performed. RESULTS Transient dark-phase hyperactivity occurred following developmental or acute exposure to PFOS or PFHxS, relative to the DMSO control. In contrast, visual startle response (VSR) hyperactivity only occurred following developmental exposure and was irreversible up to 8 dpf. Similar global transcriptomic profiles, BMCT estimates, and enriched functions were observed in PFOS- and PFHxS-exposed larvae, and ppars were identified as putative upstream regulators. Knockdown of pparda/db, but not pparaa/ab or pparg, blunted PFOS-dependent VSR hyperactivity to control levels. This finding was confirmed via antagonism of ppard in PFOS-exposed larvae. DISCUSSION This work identifies a novel adverse outcome pathway for VSR hyperactivity in larval zebrafish. We demonstrate that developmental, but not acute, exposure to PFOS triggered persistent VSR hyperactivity that required ppard function. https://doi.org/10.1289/EHP13667.

中文翻译：

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过氧化物酶体增殖物激活受体基因的研究作为暴露于结构相似的全氟烷基物质和多氟烷基物质 (PFAS) 的斑马鱼幼虫视觉惊恐反应过度活跃的要求。


背景技术全氟烷基物质和多氟烷基物质(PFAS)是在人类和环境中广泛检测到的合成化学品。此前研究表明，暴露于全氟辛烷磺酸 (PFOS) 或全氟己烷磺酸 (PFHxS) 会导致斑马鱼幼虫出现暗相过度活跃。目的 本研究的目的是阐明 PFOS 或 PFHxS 暴露导致斑马鱼幼鱼过度活跃的机制。方法 在发育期（1-4 dpf）或急性（5 dpf）暴露于 0.43-7.86μM PFOS、7.87-120μM PFHxS 或 0.4% 二甲亚砜 (DMSO) 后，对受精后 (dpf) 幼虫的游泳行为进行评估。在 4 dpf 进行发育暴露和化学清洗后，还在 5-8 dpf 评估行为。 RNA 测序用于识别全局基因表达的差异，以执行转录组基准浓度响应 (BMCT) 建模，并预测 PFOS 或 PFHxS 暴露幼虫的上游调节因子。基于 CRISPR/Cas9 的基因编辑用于在第 0 天敲低过氧化物酶体增殖物激活受体 (ppar) pparaa/ab、pparda/db 或 pparg。敲低脆片在 1-4 dpf 中暴露于 7.86μM PFOS 或 0.4% DMSO并在 5 dpf 时评估行为。还进行了与 ppard 拮抗剂 GSK3787 和 PFOS 的共暴露。结果 相对于 DMSO 对照，在发育或急性暴露于 PFOS 或 PFHxS 后出现短暂的暗相过度活跃。相比之下，视觉惊恐反应 (VSR) 过度活跃仅在发育暴露后发生，并且在 8 dpf 内不可逆转。在 PFOS 和 PFHxS 暴露的幼虫中观察到类似的整体转录组特征、BMCT 估计和丰富的功能，并且 ppars 被确定为假定的上游调节因子。 pparda/db 的敲低，但 pparaa/ab 或 pparg 的敲低，减弱了 PFOS 依赖性 VSR 过度活跃以控制水平。这一发现通过 PFOS 暴露幼虫中 ppard 的拮抗作用得到证实。讨论 这项工作确定了斑马鱼幼鱼 VSR 过度活跃的一种新的不良后果途径。我们证明，发育过程中（而非急性）接触 PFOS 会引发持续的 VSR 过度活跃，而这需要 ppard 功能。 https://doi.org/10.1289/EHP13667。