摘要
目的研究α-突触核蛋白(α-synuclein,α-Syn)对原代海马神经元膜表面N-甲基-D-天门冬氨酸(N-methyl-D-aspartate,NMDA)受体含量和功能的影响及其机制。方法细胞外添加基因重组α-Syn,使其进入原代培养神经元细胞内,观察敲减Rab5B基因前后神经元膜表面NMDA受体(NMDAR)和Rab5B表达的变化;NMDA激活NMDAR,活细胞工作站测定神经元Ca^(2+)内流变化,全细胞膜片钳记录内向跨膜电流变化。结果α-Syn增加神经元Rab5B的表达,减少膜表面NMDAR含量,并因而抑制NMDA引起的Ca^(2+)内流及内向跨膜电流;敲减Rab5B可逆转α-Syn对NMDAR的下膜作用及功能的影响。结论α-Syn通过上调Rab5B而下调海马神经元膜表面NMDAR含量及其介导的Ca^(2+)内流及内向电流。
Objective To investigate the effects of α-synuclein（α-Syn） on surface expressions and functions of N-methyl-D-aspartate（NMDA） receptors（NMDARs） in cultured primary hippocampal neurons.Methods Increased intracellular α-Syn levels were realized by extracellular addition of recombinant human α-Syn to the culture medium of rat primary hippocampal neurons.Expressions of surface NMDARs and Rab5B were observed by fluorescence immunocytochemistry and Western blotting.Rab5B antisense oligonucleotides were applied to suppress Rab5B expression.Live cell imaging system and whole-cell patch-clamp recording were used to record NMDA-evoked Ca^（2＋） influx and inward currents.Results Neurons treated with α-Syn displayed increased Rab5B,decreased surface NMDARs,and reduced NMDA-evoked Ca^（2＋） influx and inward currents.The above α-Syn effects were inhibited by suppression of Rab5B expression.Conclusion α-Syn reduces the surface expression and functions of NMDARs by increasing Rab5B expression.
出处
《首都医科大学学报》
CAS
北大核心
2017年第6期868-873,共6页
Journal of Capital Medical University
基金
国家自然科学基金(81371200,81071014,81401042)
北京市医院管理局“使命”计划专项经费资助(SML20150803)
北京市科学技术委员会资助(Z161100005116011,Z171100000117013)
北京市卫生和计划生育委员会“老年重大疾病关键技术研究”(PXM2017_026283_000002)
广西自然科学基金(2014GXNSFAA118197)
