TRPC3参与α-突触核蛋白引起的线粒体损伤

发布时间：2019-07-03 14:34

【摘要】：目的探讨经典型瞬时受体电位通道3(transient receptor potential canonical channel 3,TRPC3)是否参与α-突触核蛋白(α-synuclein,α-syn)引起的线粒体损伤。方法在α-syn过表达原代培养神经元、α-syn转基因及敲除小鼠模型,利用免疫印迹和免疫荧光技术检测TRPC3和α-syn在线粒体内的定位和表达;在原代培养神经元,运用MTT和乳酸脱氢酶法检测细胞活力和受损情况,JC-1法检测线粒体膜电势,观察敲减TRPC3对α-syn过表达引起的线粒体损伤和细胞损伤的影响。结果 TRPC3和α-syn共同表达于线粒体,过表达α-syn增加TRPC3在线粒体的分布,敲除α-syn则下调TRPC3在线粒体的分布。敲减TRPC3明显减轻过表达α-syn引起的线粒体膜电势下降和细胞毒性。结论 TRPC3可能参与过表达α-syn引起的线粒体损伤。
[Abstract]:Objective to investigate whether the typical transient receptor potential channel 3 (transient receptor potential canonical channel 3, TRPC3) is involved in the damage of mitochondria induced by 伪-synuclein, 伪-syn (伪-synaptic nucleoprotein), which is involved in the damage of mitochondria induced by 伪-synaptic nucleoprotein (伪-synaptic nuclear protein). Methods Primary cultured neurons were overexpressed in 伪-syn. 伪-syn transgenic and knockout mouse models were used to detect the localization and expression of TRPC3 and 伪-syn in mitochondria by immunoblotting and immunofluorescence. In the primary cultured neurons, the cell activity and damage were detected by MTT and lactic dehydrogenase method, and the mitochondrial membrane potential was detected by JC-1 assay. The effects of knockdown TRPC3 on the damage of mitochondria and cell injury induced by 伪-syn overexpression were observed. Results TRPC3 and 伪-syn were co-expressed in mitochondria. Overexpression of 伪-syn increased the distribution of TRPC3 in mitochondria, while knockout of 伪-syn down-regulated the distribution of TRPC3 in mitochondria. Knockout of TRPC3 significantly alleviated the decrease of mitochondrial membrane potential and cytotoxicity induced by overexpression of 伪-syn. Conclusion TRPC3 may be involved in the injury of mitochondria induced by overexpression of 伪-syn.
【作者单位】：首都医科大学基础医学院神经生物学系北京脑重大疾病研究院帕金森病研究所教育部神经变性病重点实验室;
【基金】：国家重点研究发展计划(2016YFC1306002) 国家自然科学基金(81371398,81371200) 北京市自然科学基金(7131001) 北京市创新团队建设提升计划(IDHT20140514) 北京市教育委员会科技发展计划一般项目(KM201710025001)~~
【分类号】：R-332;R742.5

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