摘要
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 effect of nov gene on differentiation of neurons and secretion of transmitter. Methods\ The nov gene was inserted into the eukaryotic expression vector pBK\|cmv in its antisense orientation between Sal I and BamH I to produce the plasmid pBK\|nov which expressed antisense RNA of nov gene by DNA recombinant technique.The plasmid pBK\|nov was transfected into cultured neurons by lipofectin co\|transfection.Positive neurons were selected in the medium supplemented with G418.The expression of endogenous nov mRNA was inhibited and measured by RT\|PCR with a pair of specific primer. Results\ The secretion of glutamine and dopamine trasmitter changed after transfected by plasmid pBK\|nov.Antisense RNA of nov gene could obviously inhibit the secretion of exiting amine trasmitter and induce the secretion of inhibiting amine trasmitter. Conclusion\ The results indicated that nov gene may play an important role in producing and secreting of neurotransmitter in the central nervous system.\;
出处
《解剖学报》
CAS
CSCD
北大核心
2001年第2期97-100,共4页
Acta Anatomica Sinica
基金
国家自然科学基金!资助项目 (3990 0 0 78)
全军 95科研基金
重庆市应用基础研究课题
关键词
反义nov基因
真核表达载体
基因转染
神经递质
Antisense nov gene
Eukaryotic expression vector
Gene transfection
Neurotransmitter
