Parkinson's disease (PD) is known as a movement disorder due to characteristic motor features. Existing therapies for PD are only symptomatic, and their efficacy decreases as disease progresses. Zebrafish, a vertebrate in which parkinsonism has been modelled, offers unique features for the identification of molecules with antiparkinsonian properties. Here, we developed a screening assay for the selection of neuroactive agents with antiparkinsonian potential. First, we performed a pharmacological validation of the phenotypes exhibited by the 6-hydroxydopamine zebrafish model, by testing the effects of known antiparkinsonian agents. These drugs were also tested for disease-modifying properties by whole mount immunohistochemistry to TH⁺ neurons and confocal microscopy in the dopaminergic diencephalic cluster of zebrafish. Next, we optimized a phenotypic screening using the 6-hydroxydopamine zebrafish model and tested 1600 FDA-approved bioactive drugs. We found that 6-hydroxydopamine-lesioned zebrafish larvae exhibit bradykinetic and dyskinetic-like behaviours that are rescued by the administration of levodopa, rasagiline, isradipine or amantadine. The rescue of dopaminergic cell loss by isradipine was also verified, through the observation of a higher number of TH⁺ neurons in 6-OHDA-lesioned zebrafish larvae treated with this compound as compared to untreated lesioned larvae. The phenotypic screening enabled us to identify several compounds previously positioned for PD, as well as, new molecules with potential antiparkinsonian properties. Among these, we selected stavudine, tapentadol and nabumetone as the most promising candidates. Our results demonstrate the functional similarities of the motor impairments exhibited by 6-hydroxydopamine-lesioned zebrafish with mammalian models of PD and with PD patients, and highlights novel molecules with antiparkinsonian potential.

由于特有的运动功能，帕金森氏病（PD）被称为运动障碍。现有的PD疗法仅是对症疗法，其功效随着疾病的进展而降低。斑马鱼是对帕金森氏症进行建模的脊椎动物，具有独特的特征，可鉴定具有反帕金森氏特性的分子。在这里，我们开发了一种筛选试验，用于选择具有抗帕金森病潜能的神经活性剂。首先，我们通过测试已知的抗帕金森病药物的作用，对6-羟基多巴胺斑马鱼模型表现出的表型进行了药理学验证。还通过针对TH ^+的整体免疫组织化学测试了这些药物的疾病缓解特性斑马鱼多巴胺能双脑簇中的神经元和共聚焦显微镜检查。接下来，我们使用6-羟基多巴胺斑马鱼模型优化了表型筛选，并测试了1600种FDA批准的生物活性药物。我们发现6-羟基多巴胺损伤的斑马鱼幼虫表现出运动迟缓和运动障碍样的行为，这些行为可以通过左旋多巴，雷沙吉兰，异地平或金刚烷胺的给药来挽救。通过观察到更高数量的TH ⁺，还证实了伊拉地平能挽救多巴胺能细胞的丢失与未经处理的病变幼虫相比，用这种化合物处理的6-OHDA病变斑马鱼幼虫中的神经元。通过表型筛选，我们可以鉴定出先前定位于PD的几种化合物，以及具有潜在抗帕金森病特性的新分子。其中，我们选择司他夫定，他喷他多和萘丁美酮作为最有前途的候选人。我们的结果证明了6-羟基多巴胺损伤的斑马鱼与PD的哺乳动物模型以及PD患者所表现出的运动障碍的功能相似性，并突出了具有抗帕金森病潜能的新型分子。