RNA干扰CRMP 5抑制大鼠海马神经元突起生长

doi:10.13418/j.issn.1001-165x.2016.03.014

中国临床解剖学杂志 ›› 2016, Vol. 34 ›› Issue (3): 303-307.doi: 10.13418/j.issn.1001-165x.2016.03.014

RNA干扰CRMP 5抑制大鼠海马神经元突起生长

李素梅¹，　李斌¹，　张吉凤^1,2，　谭明会^1,2，　武凤鸣¹，　郭国庆^1,2,3

暨南大学医学院　1.解剖学系；　2.中西医结合博士后流动站；　3.病理生理学系，广州 510632

收稿日期:2014-09-28 出版日期:2016-05-25 发布日期:2016-06-17
通讯作者: 郭国庆，教授，博士生导师，Tel：85220251，E-mail：tgqguo@jnu.edu.cn
作者简介:李素梅（1981-），女，山东人，硕士，实验师，研究方向：中枢神经的损伤与再生，Tel：85220251，E-mail：tlism@jnu.edu.cn
基金资助:
国家自然科学基金(31170941)；广东省自然科学基金项目(S2013010014191)

RNA interference with CRMP5 inhibites outgrowth of neurite in rat hippocampal neurons

LI Su-mei¹, LI Bin¹, ZHANG Ji-feng^1,2, TAN Ming-hui ^1,2, WU Feng-ming¹, GUO Guo-qing ^1,2,3

School Medicine of Jinan University 1.Department of Anatomy; 2. Post-docto ralresearch station of integrated traditional Chineseand Western Medicine; 3. Department of pathophysiology; Guangzhou 510632, China

Received:2014-09-28 Online:2016-05-25 Published:2016-06-17

摘要/Abstract

摘要：

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siRNA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 siRNA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 siRNA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。

关键词: , CRMP5, RNA干扰, 突起, 神经元, 海马

Abstract:

Objective　To investigate positive effect of CRMP5 gene on neurite outgrowth in rat hippocampal neurons.　Methods　After selecting an effective siRNA that interfered CRMP5 gene, and fluorescence radix FAM was constructed upon 5′ terminal of the siRNA. The siRNA was then transiently transfected into hippocampal neurons with lipofection reagent. The formation of neurites and branches of hippocampal neurons were observed by confocal microscopy.　Results　After the siRNAs labeled by FAM were transfected into hippocampal neurons,the FAM was distributed in the neuron cell body and neurite; Then,CRMP5 siRNA confirmed by immunofluorescence could effectively silence CRMP5 protein expression. The neurons had shorter neurites and branches than the control group after silencing CRMP5 gene expression. Compared with the control group, the length of neurites became significantly shorter after the CRMP5 siRNA was transfected into hippocampal neurons (P<0.05). No significant difference was found in the number of primary neurites, while the number of secondary and above neurites had significant difference (P<0.05).　Conclusions　Silencing of CRMP5 gene was shown to inhibit neurite growth and branch formation of hippocampal neurons.

Key words: CRMP5, RNA interference, Neurite, Neurons, Hippocampus

李素梅,李斌,张吉凤,谭明会,武凤鸣,郭国庆. RNA干扰CRMP 5抑制大鼠海马神经元突起生长[J]. 中国临床解剖学杂志, 2016, 34(3): 303-307.

LI Su-mei, LI Bin, ZHANG Ji-feng, TAN Ming-hui, WU Feng-ming, GUO Guo-qing. RNA interference with CRMP5 inhibites outgrowth of neurite in rat hippocampal neurons[J]. Chinese Journal Of Clinical Anatomy, 2016, 34(3): 303-307.

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RNA干扰CRMP 5抑制大鼠海马神经元突起生长

RNA interference with CRMP5 inhibites outgrowth of neurite in rat hippocampal neurons

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