Department of Anatomy, Hainan Medical College, Haikou 571101, Hainan, China.
Brain Res. 2011 Apr 6;1383:62-70. doi: 10.1016/j.brainres.2011.01.067. Epub 2011 Jan 26.
MicroRNAs (miRNAs) play an important role in the development, differentiation, proliferation, survival, and oncogenesis of cells and organisms including nervous system. However, the role of miRNAs in primary neurons of dorsal root ganglion (DRG) after injury was not clear. In this study, a miRNA microarray analysis was performed, and a total of 21 miRNAs were found to be down-regulated following unilateral sciatic nerve transection. The miR-144, miR-145, and miR-214 were further validated using quantitative reverse transcriptase PCR (qRT-PCR). Moreover, in situ hybridization (ISH) experiments using locked nucleic acid (LNA)-modified DNA oligonucleotide probes verified that miR-144, miR-145, and miR-214 were expressed in primary neurons of DRG and down-regulated following sciatic nerve transection. Predictions of potential miRNA targets involved were identified by performing a bioinformatics analysis. These predictions were tested using miRNA luciferase reporter vectors, with Robo2 and srGAP2 evaluated as the potential targets of miR-145 and miR-214, respectively. The role of miR-145 in cultured primary neurons was also investigated, and the result found that miR-145 miR-145 inhibited neurite growth and down-regulated Robo2 expression. Finding from this study suggested that miRNAs of DRG can mediated the course of regeneration including through Slit-Robo-srGAP signaling pathway after injury.
微小 RNA(miRNAs)在细胞和生物的发育、分化、增殖、存活和癌变中发挥重要作用,包括神经系统。然而,miRNAs 在损伤后背根神经节(DRG)原代神经元中的作用尚不清楚。在这项研究中,进行了 miRNA 微阵列分析,发现单侧坐骨神经横断后共有 21 个 miRNA 下调。使用定量逆转录 PCR(qRT-PCR)进一步验证了 miR-144、miR-145 和 miR-214。此外,使用锁定核酸(LNA)修饰的 DNA 寡核苷酸探针的原位杂交(ISH)实验证实,miR-144、miR-145 和 miR-214 在 DRG 的原代神经元中表达,并在坐骨神经横断后下调。通过进行生物信息学分析鉴定了涉及的潜在 miRNA 靶标的预测。使用 miRNA 荧光素酶报告载体测试这些预测,将 Robo2 和 srGAP2 分别评估为 miR-145 和 miR-214 的潜在靶标。还研究了 miR-145 在培养的原代神经元中的作用,结果发现 miR-145 抑制轴突生长并下调 Robo2 表达。本研究结果表明,DRG 的 miRNAs 可以通过 Slit-Robo-srGAP 信号通路介导损伤后的再生过程。