转铁蛋白受体1对小鼠海马神经元生长与分化的影响

目的探讨转铁蛋白受体1(TfR1)对小鼠海马神经元生长与分化的影响。方法取胚胎18.5 d小鼠原代海马神经元培养,培养7 d后使用TfR1 shRNA p GFP-V-RS干扰质粒转染小鼠海马原代神经元使TfR1基因沉默,采用绿色荧光蛋白(GFP)分析及Western blotting技术检测TfR1 shRNA的转染效率;4’6-二脒基-2-苯基吲哚(DAPI)及TUNEL法检测干扰后海马神经细胞凋亡情况,免疫荧光技术检测凋亡后神经细胞骨架的变化及神经元突起生长的形态特征。结果 TfR1 shRNA质粒能有效使神经细胞内TfR1基因沉默;TfR1基因沉默后,原代海马神经细胞凋亡率增加,细胞骨架部分崩解,且神经元树突突起长度明显增长(P0.05)。结论转铁蛋白受体1可通过调控小鼠海马神经元树突的生长来影响海马神经元的分化。

Effect of transferrin receptor 1 on the growth and differentiation of mouse hippocampal neurons

Objective To investigate the effect of transferrin receptor 1 (TfR1) on the growth and differentiation of mouse hippocampal neurons.Methods Hippocampal neurons were isolated from C57BL/6 mouse at embryonic day 18.5,and then transfected by TfR1 shRNA pGFP-V-RS plasmid after 7 days culture.The transfection efficiency of TfR1 shRNA was detected by green fluorescent protein (GFP) and Western blotting ting assay.4-6-diamidino-2-phenylindole (DAPI) and TUNEL stainings were used to detect the apoptosis of hippocampal neurons.Immunofluorescence staining was used to analyze the changes of ncuronal cytoskeleton and the morphological characteristics of neurite growth.Results TfR1 shRNA interference plasmid effectively knockdowned TfR1 gene in cultured neurons.TfR1 shRNA interference obviously induced neurons apoptosis and the disintegration of cytoskeletons.Accordingly,the neurite elongation significantly increased after TfR1 gene interfered (P ＜ 0.05).Conclusion TfR1 can affect the differentiation of hippocampal neurons by regulating the neurite outgrowth of mouse hippocampal neurons.