中国神经再生研究(英文版) ›› 2014, Vol. 9 ›› Issue (10): 1031-1040.doi: 10.4103/1673-5374.133164

• 原著:周围神经损伤修复保护与再生 • 上一篇    下一篇

影响坐骨神经损伤后再生的背根神经节差异表达基因

  

  • 收稿日期:2014-04-02 出版日期:2014-05-26 发布日期:2014-05-26
  • 基金资助:

    中国国家自然科学基金项目(81160158,30860290)

Differential expression of microRNAs in dorsal root ganglia after sciatic nerve injury

Anjie Lu 1, Zufa Huang 1, Chaoyue Zhang 1, Xianfang Zhang 2, Jiuhong Zhao 2, Haiying Zhang 2, Quanpeng Zhang 2, Song Wu 1, Xinan Yi 2   

  1. 1 Department of Orthopedics, the Third Xiangya Hospital, Central South University, Changsha, Hunan Province, China
    2 Neurosciences Research Department, Hainan Medical University, Haikou, Hainan Province, China
  • Received:2014-04-02 Online:2014-05-26 Published:2014-05-26
  • Contact: Xinan Yi, M.D., Neurosciences Research Department, Hainan Medical University, Haikou 571101, Hainan Province, China, xinanyi2011@yahoo.cn.
  • Supported by:

    This work was supported by the National Natural Science Foundation of China, No. 81160158 and 30860290.

摘要:

实验旨在了解通过调节srGAP3表达参与周围神经再生过程的微小RNA。首先通过微阵列芯片分析筛选出坐骨神经切断后背根神经节中差异表达的微小RNA,共发现23种微小RNA在坐骨神经切断后背根神经节中表达变化显著。之后通过生物信息学分析确定微小RNA-214以srGAP3为可能靶基因,且与轴突导向信号通路相关性最大;并以qRT-PCR、双荧光素酶报告基因方法分别证实了微小RNA-214的下调及其与srGAP3的靶向关系。在坐骨神经损伤后1-7 d,核酸原位杂交、western blot和免疫荧光染色发现,大鼠背根神经节中微小RNA-214表达逐渐下降,而srGAP3蛋白表达逐渐增高。可见坐骨神经损伤可诱导腰段背根神经节中多种微小RNA的差异表达,其中微小RNA-214可能与srGAP3存在靶向关系,且二者表达呈反变关系。
 

关键词: 神经再生, 周围神经损伤, 坐骨神经损伤, Slit-Robo GTP酶激活蛋白3, 微小RNA-214, 背根神经节, 基因表达, 基因芯片, 生物信息学

Abstract:

This study investigated the possible involvement of microRNAs in the regulation of genes that participate in peripheral neural regeneration. A microRNA microarray analysis was conducted and 23 microRNAs were identified whose expression was significantly changed in rat dorsal root ganglia after sciatic nerve transection. The expression of one of the downregulated microRNAs, microRNA-214, was validated using quantitative reverse transcriptase-PCR. MicroRNA-214 was predicted to target the 3′-untranslated region of Slit-Robo GTPase-activating protein 3. In situ hybridization verified that microRNA-214 was located in the cytoplasm of dorsal root ganglia primary neurons and was downregulated following sciatic nerve transection. Moreover, a combination of in situ hybridization and immunohistochemistry revealed that microRNA-214 and Slit-Robo GTPase-activating protein 3 were co-localized in dorsal root ganglion primary neurons. Western blot analysis suggested that Slit-Robo GTPase-activating protein 3 was upregulated in dorsal root ganglion neurons after sciatic nerve transection. These data demonstrate that microRNA-214 is located and differentially expressed in dorsal root ganglion primary neurons and may participate in regulating the gene expression of Slit-Robo GTPase-activating protein 3 after sciatic nerve transection.

Key words: nerve regeneration, peripheral nerve injury, sciatic nerve injury, Slit-Robo GTPase-activating protein 3, microRNA-214, dorsal root ganglia, gene expression, microarray, bioinformatics, NSFC grant, neural regeneration