应用锚定消减杂交技术克隆分析大鼠脊髓原代神经元损伤、修复相关基因

Rat spinal cord primary neuron injury-regeneration-related genes cloned by improved subtractive hybridization

目的克隆大鼠脊髓原代神经元细胞损伤、修复相关基因,从分子水平探讨中枢神经系统损伤修复的机制。方法建立脊髓原代神经元体外损伤模型,运用改良的锚定消减杂交技术,克隆分析与原代脊髓神经元细胞损伤、修复相关的基因。结果随机挑选110个消减杂交得到的cDNA克隆,测序分析得到27个有功能线索的差异表达序列,其中23个为已知基因,包括Snc、Clu等与神经系统发育及退行性病变相关基因。结论运用锚定消减杂交技术成功克隆了一批与脊髓神经元损伤、修复相关的基因,为探求中枢神经系统损伤修复的分子机制奠定了基础。

Objective To clone injury regeneration related genes after primary neurons of spinal cord were injured in rat, and to elucidate the molecular mechanism of injury and regeneration in CNS. Methods An in vitro spinal cord primary neuron injury model was replicared in rat. The differentially expressed genes during injury and regeneration periods of spinal cord primary neurons were cloned by improved subtractive hybridization. Results Twenty seven differentially expressed sequences were obtained, 23 were known genes and the others were unknown. Among these known sequences, cDNA for synuclein and clusterin, together with several other gene sequences, were closely related with nerve degenerative diseases. Conclusion Primary neuron injury-regeneration-related genes of the spinal cord can be successfully cloned by improved subtractive hybridization.