Zebrafish Assay for Parkinson's Disease Drugs

描述（由申请人提供）：描述：本研究计划的目标是开发一种快速的体内试验，用于筛选小分子在整个脊椎动物生物体中保护或再生神经元的能力。在第一阶段，斑马鱼胚胎中的多巴胺能神经元，通过转基因掺入荧光蛋白而变得容易看到，将暴露于神经毒素MPTP。将基于神经元荧光的变化来测量神经元的损伤。然后将测试已知功能的分子以及新型分子以剂量依赖性方式保护神经元或促进神经元生长的能力。通过使用高含量荧光成像仪来量化96或384孔板格式中的荧光变化，可以加速这一奋进的成功，以允许在II期中进行更高通量的筛选。在该系统中鉴定的分子具有作为治疗神经退行性疾病如帕金森病的新型药物的潜力。目前没有任何治疗方法可以阻止帕金森病神经细胞死亡的进展，这增加了该领域药物发现创新的需求。由于目前利用培养中的细胞或外植体的高通量技术可能无法反映真实的体内环境，因此斑马鱼测定将代表公共卫生研究的化合物筛选的重大创新进展：本项目的目标是开发荧光斑马鱼测定来监测神经毒素诱导的神经元损伤。保护神经元免受损伤的药物可能对治疗帕金森病有用。

DESCRIPTION (provided by applicant): Description: The goal of this research program is to develop a rapid, in vivo assay for screening small molecules for their ability to protect or regenerate neurons in a whole vertebrate organism. In Phase I, dopaminergic neurons in zebrafish embryos, made easily visible by transgenic incorporation of a fluorescent protein, will be exposed to the neurotoxin MPTP. Damage to neurons will be measured based on changes in fluorescence of the neurons. Molecules of known function, as well as novel molecules, will then be tested for their ability to protect neurons or promote neuronal growth in a dose-dependent fashion. Success in this endeavor can be accelerated to allow higher throughput screening in Phase II by using a high content fluorescent imager to quantify changes in fluorescence in a 96 or 384-well plate format. Molecules identified in this system have the potential to serve as novel drugs for the treatment of neurodegenerative diseases such as Parkinson's disease. No current treatments have been shown to prevent the progression of neuronal cell death in Parkinson's disease, increasing the need for innovation in drug discovery in this area. Since current high throughput technologies which utilize cells or explants in culture may not reflect the true in vivo environment, the zebrafish assay will represent a significant innovative advance in compound screening of research to public health: The goal of this project is develop a fluorescent zebrafish assay to monitor damage to neurons induced by neurotoxins. Drugs that protect neurons from damage may be useful for the treatment of Parkinson's disease.