为了进一步明确METH作用后神经退行性变标志蛋白α-SN与 tau 蛋白的相互关系及在神经毒性机制中所起的协同作用，在前期工作基础上，我们拟利用基因敲除和RNAi技术完全和非完全性去除其中一种蛋白的影响，在体内、外水平上利用分子生物学技术、分子病理技术、细胞生物学技术和动物行为学等技术研究METH作用后观察二者总表达量、寡聚体、磷酸化、硝基化及其相关因子的改变，观察二者分布、共定位表达及复合寡聚体的变化情况，探究其相互作用对氧化应激、线粒体损伤、内质网应激、细胞凋亡和突触重塑性等神经毒性损伤相关指标以及动物行为学的影响，阐明其协同作用在上述神经毒性机制中的重要功能，初步寻找其在相邻神经元间传播的佐证。希望通过本研究能够更深入了解α-SN和tau相互作用在METH神经毒性分子机制中所扮演的重要角色，为阐明其关键靶点作用和戒毒解毒研究提供基础，也为相关法医学分子诊断和毒理机制探讨提供参考。

In order to clarify the interaction and synergistic effect between α-synuclein and tau protein in METH-induced neurotoxicity, we plan to use gene knockout and RNAi technology to regulate the expression level of target protein in vitro and in vivo in this study. Based on our established molecular biological technique, molecular pathology, cell biological technique, ethology and so on, we will detect the total α-synuclein and tau protein, oligomers, protein phosphorylation, nitration and other related factors with or without METH treatment, observe their distribution, colocalization and composite-oligomers,and explore the effects on neurotoxic-related indicators including oxidative stress, mitochondrial damage, endoplasmic reticulum stress, apoptosis and synaptic remodeling and animal behavior caused by this interaction. We will also search the possible evidence of this two proteins spreading between neighboring neurons. This project will be helpful to clarify the molecular mechanism of α-synuclein and tau interaction in METH-induced neurotoxicity, and provide a richer theoretical basis for the therapy of drug addiction as well as the references for the related forensic molecular diagnosis and forensic toxicological mechanism.