新生大鼠脊髓损伤修复差异表达基因的筛选

Screening of differentially expressed genes in the regeneration of injured spinal cord of neonatal rats

目的 探讨新生大鼠半横切后再生时运动皮层神经元基因表达变化及其机制。方法 新生大鼠脊髓单侧横断后即刻局部接受胚胎脊髓移植 ,术后 3d用抑制性消减杂交技术筛选大鼠双侧运动皮层神经元差异表达基因 ,构建差减cD NA文库 ,通过蓝白斑实验和RNA斑点杂交验证差异表达基因真阳性克隆 ,测序分析。结果 在Genbank中检索出 11个上调表达的基因分别与 11个不同的蛋白基因有明显同源性 ,7个下调表达的基因分别与 7个不同的蛋白基因有明显同源性。结论 新生大鼠运动皮层神经元轴突中断后的再生过程中有基因表达变化 。

Objective To identify the differentially expressed genes of the regenerating motor cortical neurons after hemi section of the corticospinal tract (CST) of the newborn rats so as to investigate the mechanism in regeneration. Methods After hemi section of the unilateral CST of the newborn rats, fetal spinal cord tissue was transplanted immediately at the local site of the lesion. The differentially expressed genes of bilateral motor cortical neurons of the newborn rats were screened at 3 d after operation using suppression subtractive hybridization method for the construction of differential expression library. The really positive clones were confirmed by blue/white spot test and dot hybridization technique, and their sequences were analyzed. Results Compared with the published expressed sequence tags in GenBank, we found that 11 up regulated genes and 7 down regulated genes showed distinct homology with 11 and 7 different protein genes, respectively. Conclusion There are gene expression changes in the regenerating process of the motor cortical neurons of the newborn rats after fetal spinal cord transplantation. The differential genes may be associated with the regeneration ability of motor cortical neurons.