突触囊泡蛋白synaptotagmin XI的过表达对PC12细胞和聚集小体形成的影响  

Effects of Synaptotagmin XI Plasmid Overexpression on PC12 Cells and the Aggresomes Formation

作　　者：郑汭[1] 王志全[1] 盛呈雨[1] 周海燕[1] 丁健青[1] 陈生弟[1] 

机构地区：[1]上海交通大学医学院附属瑞金医院神经科,上海交通大学医学院神经病学研究所,上海200025

出　　处：《中国神经免疫学和神经病学杂志》2007年第5期304-306,316,I0002-I0003,共6页Chinese Journal of Neuroimmunology and Neurology

基　　金：国家重点基础研究发展计划(973计划;2006CB500706);上海市医学领军人才资助项目(LJ06003)

摘　　要：目的构建突触囊泡蛋白synaptotagmin XI(SytXI)重组质粒,观察其在PC12细胞中的表达,以及其对细胞和聚集小体形成的影响。方法运用人胚脑的cDNA文库作为模板,pcDNA3.1myc-His(-)B质粒作为载体,运用基因克隆技术构建重组质粒,限制性内切酶酶切,测序鉴定重组质粒;运用脂质体将重组质粒导入PC12细胞,Annexin V-PI双染色和免疫荧光观察对细胞的影响。结果测序鉴定质粒构建成功,过表达SytXI导致细胞死亡,增加α-synuclein和vimentin阳性聚集小体的形成。结论成功构建SytXI重组表达质粒,过表达造成对PC12细胞的毒性作用,并促进聚集小体的形成。Objective To investigate the construction of synaptotagmin XI （SytXI） plasmid and overexpression, and its impact on PC12 cells and the aggresome formation. Methods SytXI was obtained by using human fetal brain cDNA library as template and then ligated into pcDNA3, lmyc-His（-）B plasmid. The recombinant plasmid was identified by endonuclease and DNA sequencing. After transfected into PC12 cells, Annexin-V-PI staining and immunofluorescence were used to define its impact on the cells. Results The SytXI gene was ligated correctly into the vector. Overexpression of SytXI led to cell necrosis, and increased the formation of a-synuclein and vimentin positive aggresomes. Conclusions The recombinant plasmid was constructed and successfully transfected into PC12 cells. The overexpression of SytXI was toxic to PC12 cells and increased the formation of aggresomes.

关 键 词：SYNAPTOTAGMIN Ⅺ 聚集小体 帕金森病 

分 类 号：R742.5[医药卫生—神经病学与精神病学]