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目的 研究 nov基因在神经元功能分化和神经递质分泌中的作用。 方法 利用反义核酸技术构建成反义 nov基因真核表达载体 p BK- nov。采用脂质体转染技术 ,将反义表达载体导入培养的大鼠大脑皮质神经元和纹状体神经元 ,经 G418筛选出被转染的神经元继续培养。RT- PCR检测转染后 nov m RNA的表达量。采用高效氨基酸分析仪和高效液相色谱仪 ,检测抑制 nov基因表达后氨基酸类递质和多巴胺类递质的变化。 结果 谷氨酸和肾上腺素的分泌量明显下降 ,γ-氨基丁酸上升 ,而对多巴胺的分泌没有显著的影响。 结论 以上结果提示 ,nov基因在中枢神经系统神经递质的生成释放和代谢中起作用 ,可能籍此调节神经系统的功能。
Objective To study the role of nov gene in neuronal differentiation and neurotransmitter secretion. Methods Antisense oligonucleotides were used to construct pBK-nov, an antisense nov gene eukaryotic expression vector. The antisense expression vector was transfected into cultured rat cortical neurons and striatum neurons by liposome transfection technique. The transfected neurons were screened by G418 for further culture. The expression level of nov m RNA after transfection was detected by RT-PCR. High-performance amino acid analyzer and high-performance liquid chromatography were used to detect the changes of amino acid neurotransmitters and dopamine neurotransmitters after inhibiting the expression of nov gene. The results of glutamate and epinephrine secretion decreased significantly, γ-aminobutyric acid increased, while the secretion of dopamine no significant impact. Conclusions The above results suggest that the nov gene may play a role in the production, release and metabolism of neurotransmitters in the central nervous system and may thus regulate the function of the nervous system.