Ⅱ型单胺囊泡转移体在SH-SY5Y细胞中的稳定表达

Stable Expression of Vesicular Monoamine Transporter 2 in SH-SY5Y Cells

目的构建稳定表达Ⅱ型单胺囊泡转移体(VMAT2)的SH-SY5Y细胞系。方法构建VMAT2真核表达载体,利用脂质体转染神经母细胞瘤SH-SY5Y,选择筛选后挑取单克隆扩大培养,通过Western-blot及免疫荧光的方法鉴定蛋白质在细胞中的表达,并通过免疫双荧光技术对VMAT2在细胞系中的分布进行定位。结果Western-blot及免疫荧光结果显示该蛋白在挑选的细胞系中有高效表达。免疫双荧光结果显示表达的蛋白定位在大密度核心囊泡(LDCV)。结论该细胞系的成功构建为研究VMAT2在帕金森氏病(PD)发病机制中的作用奠定良好的基础。

Objective To establish the SH-SYSY cell line stably expressing vesicular monoamine transporter 2 （VMAT2）. Methods An eukaryotic expression vector of VMAT2 was constructed and then transfected into SH-SYSY cells by Lipofectamine. The expression of VMAT2 in SH-SYSY cell line was detected by Western-blot and immunofluorescence technique. Double immunofluorescence technique was used to determine the localization of VMAT2 in the cells. Results VMAT2 was highly expressed in SH-SYSY cell line revealed by Western-blot and immunofluorescence and was found to locate on LDCV shown by double immunofluorescence technique. Conclusion This work has laid a good foundation for the further studies on the role of VMAT2 in Parkinson＇ s Disease （ PD）.