Dok 6促进神经营养因子3诱导的过表达原肌球蛋白相关激酶C的PC12细胞突起生长

Dok 6 Promote Neurite Outgrowth of Tropomyosin-related Kinase C Overexpressed PC12 Cells in Stimulation of Neurotrophin-3

目的探讨胞浆接头蛋白Dok6对PC12细胞突起生长的影响。方法将Dok6的不同功能模体片段构建至TrkC/Y516F突变体中形成融合克隆。利用Western blot法验证各个融合克隆在细胞内是否稳定表达。利用瞬时转染的方法将融合克隆导入PC12细胞进行过表达,之后用NT-3刺激PC12细胞突起生长,检测不同Dok6片段的效应。结果各个融合克隆均能在细胞内稳定表达。融合了Dok6 PTB结构域+C末端以及单纯C末端的融合克隆均可以挽救由于TrkC受体516位酪氨酸突变导致的PC12细胞突起生长减少,而融合单纯的Dok6 PTB结构域则无此效应。结论Dok6可以促进NT-3诱导的过表达TrkC的PC12细胞突起生长,并且此效应与其C末端密切相关。

Objective To study the role of adaptor protein Dok 6 in neurite outgrowth in PC12 cells.Methods Series of fusion clones were constructed by fusing different domains of Dok 6 into mutant TrkC/Y516F.These constructs were transiently transfected into PC12 cells separately and the expression levels of fusion proteins were detected by Western blot.Neurite outgrowth in these PC12 cells was tested after stimulation of NT-3.Results Each fusion clone was stably expressed in PC12 cells.The fusion clones that fused bo...