样品富集-毛细管电泳-快速循环伏安研究帕金森病模型果蝇神经系统

Parkinson's disease(PD) is the second most common neurodegenerative disease; however, little is known about the PD mechanism. In our study, Drosophila would be chosen as animal model of Parkinson's disease. The amount of neurotransmitters and metabolites in the brain of wild type and Genetic mutant Drosophila are monitored by capillary electrophoresis fast-scan cyclic voltammetry during development. The influence of environment, Neurotoxin and genetic mutant would be tested. The difficult in analyzing neurotransmitters and metabolites in single brain of Drosophila lies in the small amount of analytes (fmol~amol)and the complex sample matrix. Sample pre-concentration and capillary electrophoresis on-line enrichment would be adopted to improve the sensitivity of the method. The developed method would be used to monitor the amount of neurotransmitters and metabolites in the brain of wild type Drosophila, Drosophila Neurotoxin Models of PD, and Drosophila genetic mutant models of PD. The research findings will enrich the knowledge of neuron development and provide more information about the PD mechanism. At the same time, the system developed can be used in drug screening for PD.

帕金森病是第二大类神经退行性疾病，尚不清楚其发病机制。本项目以果蝇为帕金森病模型动物，通过监测其成长过程中药物、转基因等对神经递质及代谢物含量的影响，探究帕金森病的发病机理。针对单个果蝇大脑体积小（<8纳升）、神经递质含量低（飞~阿摩尔数量级）、样品基质复杂的特点，为检测单个果蝇脑内的神经递质及其代谢物，我们将开展以下研究：研制小体积样品的进样装置；借助"样品前处理"和"毛细管电泳在线富集"预富集待测物；采用"毛细管电泳-快速循环伏安法"选择性检测该类物质。该方法用于监测"药物诱导帕金森病模型"和"转基因帕金森病模型"果蝇的脑内神经递质及代谢物随时间的变化，绘制神经递质及代谢物的时间曲线，通过与野生果蝇及转基因果蝇原种的成长曲线对比，寻找神经系统发育过程中各种因素的影响规律。该项目的研究成果有助于：揭示脑内神经递质及代谢物等随时间的变化规律；探究帕金森病的发病机理；筛选治疗帕金森病的药物。

本项目以果蝇为帕金森模型生物,通过检测其成长过程中年龄、转基因等因素对神经递质及其代谢物含量的影响，探究帕金森疾病的发病机理。我们提出＂压力辅助-场放大进样-毛细管电泳-快速循环伏安＂技术，实现了果蝇中枢神经系统内痕量神经递质及其代谢物的同时检测。我们测定了＂野生型＂果蝇与＂转基因帕金森病模型＂果蝇中枢神经系统内神经递质及代谢物含量随时间变化的情况，绘制了神经递质的时间变化曲线。我们发现，与参照组相比，帕金森疾病模型果蝇中枢神经系统内多巴胺的含量在晚期出现减少趋势，这与文献报道的帕金森疾病患者脑内多巴胺神经元的衰退相符；同时，色氨酸和真鞘胺的含量却显著增加。这些结果有助于我们更深入的了解神经系统的发育规律，为更好的了解帕金森疾病的发病机理提供帮助。