基于多巴胺能通路探讨新型全氟化合物OBS诱导斑马鱼神经行为毒性效应与分子机制

近年来，全氟壬烯氧基苯磺酸钠（OBS）在不同环境介质中的存在和在食物链中的生物富集与放大，引起人们对其潜在生物学毒性效应的关注。我们近期研究发现高浓度OBS可通过干扰多巴胺分泌诱发斑马鱼急性神经行为毒性，纤毛形成障碍可能引发了多巴胺分泌的失衡。然而，环境浓度下OBS神经行为毒性效应及纤毛介导的多巴胺分泌失调分子机制未知。本项目拟以斑马鱼为模式生物，以多巴胺能通路为切入点，开展环境浓度下OBS神经行为毒性效应评估；并结合iTRAQ技术，对暴露后纤毛发生相关差异表达基因进行Morphlino敲降或mRNA过表达，通过观察回补实验中多巴胺能表型是否回复确定破坏多巴胺分泌的关键因子，利用WISH和Pull Down技术，阐明OBS通过作用于纤毛发生关键因子破坏纤毛形成干扰多巴胺分泌诱导神经行为毒性，揭示OBS神经行为毒性分子机理，为OBS环境健康风险评估和毒性生物标记物建立提供理论依据。

In recent years, the existence of sodium p-perfluorous nonenoxybenzene sulfonate (OBS) in different environmental matrices, as well as its bioaccumulation and biomagnification in food chain, arouses people’s attention to its potential biological toxic effects. Our recent research found that OBS of high concentration led to acute neurobehavioral toxicity to zebrafish through interfering with dopamine secretion. Ciliary dysfunction may give rise to the abnormal dopamine secretion. However, the neurobehavioral toxicity of OBS at environment related concentrations and the molecular mechanism of cilia mediated abnormal dopamine secretion are still unknown. This study will utilize zebrafish as a model organism to evaluate the neurobehavioral toxic effects of OBS under environmental concentrations based on dopaminergic pathway, and combine with iTRAQ technology to search for the key ciliogenesis related factor destroying dopamine secretion through observing whether the dopaminergic phenotypes are rescued after knocking down by morphlino or overexpressing mRNA of cilia related differential expression genes. We will also take advantage of WISH and Pull Down to elucidate that OBS acts on key factor to disrupt ciliary formation, interfering with dopamine secretion and finally causes neurobehavioral toxicity. It will reveal the molecular mechanism of OBS induced neurobehavioral toxicity, providing the theoretical basis for environmental health risk assessment and establishment of toxic biomarker.