项目名称: 狂犬病病毒街毒株感染导致神经元树突棘变性的分子机制
项目编号: No.31272579
项目类型: 面上项目
立项/批准年度: 2013
项目学科: 农业科学
项目作者: 张茂林
作者单位: 吉林大学
项目金额: 84万元
中文摘要: 树突棘为神经元树突表面的突起物,充当神经兴奋性突触的突触后膜。本项目前期工作表明,狂犬病病毒街毒株(SRV)可引起树突棘变性,但其机制并不清楚。为此,本研究拟应用融合荧光蛋白的骨架蛋白质粒转染原代海马神经元细胞,观察SRV感染后细胞骨架的变化,同时应用骨架形成调节剂,确定在SRV感染及树突棘变性过程中涉及的主要骨架类型;应用基因表达谱芯片及定量 PCR等筛选SRV感染发生差异表达的主要骨架蛋白;将SRV结构蛋白质粒转染的神经元骨架变化与SRV感染的比较,确定主要结构蛋白,并通过与结构蛋白相互作用研究,从中筛选出关键骨架蛋白;在体内对关键骨架蛋白调控表达,结合SRV感染,检测其对树突棘及SRV增殖的影响;将所得结果在体内验证,推断SRV感染引起树突棘变性的机制。本研究首次通过树突棘骨架变化探索SRV致神经功能异常的机制,研究结果还可用于SRV逆轴浆运输及跨突触传播机制的阐释。
中文关键词: 狂犬病病毒;树突棘;微丝;微管;神经功能异常
英文摘要: Neurons are polarized with axons and dendrites.Dendritic spines,which are small protrusions of various shapes on a dendrite and the postsynaptic element of most excitatory synapses,play pivotal roles in nerve functions.Our previous work demonstrated that dedritic degeneration could be shown in neurons after street rabies viruses(SRV) infection.However,the mechanisms are not clear.In this proposal,primary hippocampus neurons will be prepared and transfected by fluorescent-fused actin or globulin plasmids, then the transfected neurons are infected with SRV and the cytoskeleton changes will be confirmed.Cytoskeleton formation inhibitors also will be used to identify the main transport processes between actin and microtubule involving in SRV infection and dedritic spines morphology.Based on above observation, the differential expressed cytoskeleton proteins and the main structural protein via SRV infection will be screened.The key skeleton proteins were selected by the interaction between the main structural protein of SRV and the differential expressed cytoskelton preoteins.The key skeleton proteins will be regulated by siRNA or high expression plasmids transfection in vitro,and their functions will be investigated in cytoskeleton formation、spine morphology and SRV multiplication after SRV infection. We will fin
英文关键词: rabies virus(RV);dendritic spine;actin;microtubule;neuronal dysfunction