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目的探讨CRMP5对大鼠海马神经元突起生长的影响。方法将带FAM标记的CRMP5的特异性干扰片段及阴性对照转染培养成熟的海马神经元,用免疫荧光的方法验证干扰片段对神经元内源性CRMP5的干扰效果,并利用共聚焦显微镜观察神经元突起以及侧枝的形成。结果携带FAM的si RNA可以成功的进入细胞,分布于神经元的胞体以及树突;免疫荧光证实CRMP5 si RNA可以有效的沉默CRMP5蛋白的表达;沉默CRMP5基因表达后的海马神经元突起短小,而且缺少分支,而对照细胞突起长,分支多;定量分析显示,导入CRMP5 si RNA的细胞突起的长度较对照细胞缩短,差异显著(P<0.05);突起的数目比较,一级突起数目无显著差异,而二级及其以上突起的数目明显减少,差异显著(P<0.05)。结论沉默CRMP5可抑制海马神经元突起的生长和侧枝形成。
Objective To investigate the effect of CRMP5 on neurite outgrowth in rat hippocampus. Methods The specific interfering fragments of CRMP5 with FAM labeling and the negative control were transfected into mature hippocampal neurons. The interference effect of the interfering fragments on endogenous CRMP5 in neurons was verified by immunofluorescence. The neurons were observed by confocal microscopy Protuberances and the formation of collateral. Results The si RNAs carrying FAM could successfully enter the cells and distributed in the soma and dendrites of neurons. The immunofluorescence confirmed that CRMP5 si RNA could effectively silence the expression of CRMP5 protein. The silenced hippocampal neurons after CRMP5 gene expression were short, The number of protuberances of CRMP5 si RNAs was shorter than that of control cells (P <0.05). The number of protuberances was not significantly different between the number of protuberances , While the number of secondary and above processes significantly reduced, the difference was significant (P <0.05). Conclusion Silencing CRMP5 inhibits neurite outgrowth and collateral formation in hippocampus.