Brenet Alexandre, Somkhit Julie, Hassan-Abdi Rahma, Yanicostas Constantin, Romain Christiane, Bar Olivier, Igert Alexandre, Saurat Dominique, Taudon Nicolas, Dal-Bo Gregory, Nachon Florian, Dupuis Nina, Soussi-Yanicostas Nadia
NeuroDiderot, Inserm, Université de Paris, 75019, Paris, France.
Département de toxicologie et risques chimiques, Institut de Recherche Biomédicale des Armées (IRBA), 91 220, Brétigny-sur-Orge, France.
Sci Rep. 2020 Nov 5;10(1):19228. doi: 10.1038/s41598-020-76056-8.
With millions of intoxications each year and over 200,000 deaths, organophosphorus (OP) compounds are an important public health issue worldwide. OP poisoning induces cholinergic syndrome, with respiratory distress, hypertension, and neuron damage that may lead to epileptic seizures and permanent cognitive deficits. Existing countermeasures are lifesaving but do not prevent long-lasting neuronal comorbidities, emphasizing the urgent need for animal models to better understand OP neurotoxicity and identify novel antidotes. Here, using diisopropylfluorophosphate (DFP), a prototypic and moderately toxic OP, combined with zebrafish larvae, we first showed that DFP poisoning caused major acetylcholinesterase inhibition, resulting in paralysis and CNS neuron hyperactivation, as indicated by increased neuronal calcium transients and overexpression of the immediate early genes fosab, junBa, npas4b, and atf3. In addition to these epileptiform seizure-like events, DFP-exposed larvae showed increased neuronal apoptosis, which were both partially alleviated by diazepam treatment, suggesting a causal link between neuronal hyperexcitation and cell death. Last, DFP poisoning induced an altered balance of glutamatergic/GABAergic synaptic activity with increased NR2B-NMDA receptor accumulation combined with decreased GAD65/67 and gephyrin protein accumulation. The zebrafish DFP model presented here thus provides important novel insights into the pathophysiology of OP intoxication, making it a promising model to identify novel antidotes.
每年有数百万起中毒事件,导致超过20万人死亡,有机磷(OP)化合物是全球重要的公共卫生问题。OP中毒会引发胆碱能综合征,伴有呼吸窘迫、高血压和神经元损伤,可能导致癫痫发作和永久性认知缺陷。现有的应对措施能挽救生命,但无法预防长期的神经元合并症,这凸显了迫切需要动物模型来更好地理解OP神经毒性并确定新型解毒剂。在此,我们使用二异丙基氟磷酸酯(DFP),一种典型的中度毒性OP,结合斑马鱼幼体,首次表明DFP中毒导致主要的乙酰胆碱酯酶抑制,引发麻痹和中枢神经系统神经元过度激活,表现为神经元钙瞬变增加以及即刻早期基因fosab、junBa、npas4b和atf3的过表达。除了这些癫痫样发作事件外,暴露于DFP的幼体显示神经元凋亡增加,地西泮治疗可部分缓解这两种情况,表明神经元过度兴奋与细胞死亡之间存在因果关系。最后,DFP中毒导致谷氨酸能/γ-氨基丁酸能突触活动平衡改变,NR2B-NMDA受体积累增加,同时GAD65/67和gephyrin蛋白积累减少。本文提出的斑马鱼DFP模型因此为OP中毒的病理生理学提供了重要的新见解,使其成为确定新型解毒剂的有前景的模型。