摘要
目的建立α-突触核蛋白(α-synuclein,α-syn)聚集及传播的细胞和动物模型,为帕金森病的发病机制研究提供基础。方法亲和层析法纯化α-syn蛋白,体外诱导其聚集成为α-syn纤维(Preformed fibrils,PFFs);培养稳定表达GFP-α-syn的HEK293细胞系及原代神经元,转导α-synPFFs后免疫荧光染色法观察细胞内α-syn聚集情况;小鼠立体定位注射α-syn PFFs,免疫组织化学法检测内源性α-syn的聚集及传播情况。结果纯化的α-syn可在体外聚集形成聚集体;在细胞及动物水平观察到α-syn PFFs可诱导内源性蛋白的聚集和传播。结论本研究建立了α-syn聚集及传播的细胞和动物模型,为帕金森病的相关研究打下了基础。
Objective To establish the cell and animal models of α-synuclein aggregation and propagation for the study of Parkinson’s disease.Methods Recombinant α-synuclein(α-syn) was purified by affinity chromatography and aggregated into α-synuclein preformed fibrils(α-syn PFFs) in vitro. HEK293 cell line expressing GFP-α-synuclein and primary neurons were transduced with α-syn PFFs, immunofluorescence staining were used to detect endogenous protein aggregation. The accumulation and propagation of endogenous α-syn were detected by immunohistochemistry after stereotactic injection of α-syn PFFs into mice brain.Results Purified recombinant α-syn aggregated into PFFs in vitro. α-syn PFFs induced the aggregation and propagation of endogenous protein both in cell and mouse models.Conclusion The cell and animal models of α-syn aggregation and propagation were established both in vitro and in vivo. This would lay a foundation for further study of Parkinson’s disease.
作者
潘丽娜
熊敏
田野
李春蕊
张振涛
Pan Lina;Xiong Min;Tian Ye(Department of Neurology,Renmin Hospital of Wuhan University,Wuhan 430060)
出处
《卒中与神经疾病》
2020年第6期701-704,共4页
Stroke and Nervous Diseases
基金
国家自然科学基金(No.81822016,81771382)。
