摘要
目的检测小鼠胚胎干细胞(ES)体外向神经元诱导分化过程中,7种重要时钟基因的表达情况。方法小鼠胚胎十细胞(ES),经过5个阶段诱导分化为神经元。各阶段细胞的总 RNA反转录为 cDNA,利用7种时钟基因的特异引物进行 PCR 反应,以明确其随分化表达的情况。结果各个时钟基因无非特异扩增,BMAL1、CLOCK、CRY1和 CRY2、PER1、PER3基因在各个分化阶段中都表达,而 PER2基因只在终末阶段被检测到。结论时钟基因的转录在 ES 细胞向神经元分化的过程中并不一致,提示在成体动物存在的时钟基因环路,在 ES 细胞向神经元分化过程中还没有建立。PER2 仅在分化终末期——大量停止分裂的神经元和胶质细胞出现的时期才转录,提示其可能在细胞增殖过程中发挥某种作用。
Objective To analysis the expression profile of 7 clock genes during mouse embryonic stem cell （mES） differentiation. Methods Mouse ES cells of the line 129 were cultured and induced to differentiate into neurons by 5 stages method. The expression of 7 clock genes ： BMAL1, CLOCK, CRY1, CRY2,PERI, PER2, and PER3 in the five stages were determined by RT-PCR. Results In the amplification condition, no non-specific band was found, and only bands of expected sizes were detected. Six clock genes,BMAL1, CLOCK, CRY1, CRY2,PER1 and PER3 were expressed in 129 ceils at all five stages in the differentiation process, however, PER2, could only be determined in the last stage. Conclusion The transcription profile of the clock genes is different during the differentiation of the ES cells towards neurons. The transcription of the gene PER2 is limited to the final stage, when postmitotic neurons and astrocytes emerged. A clock genes loop existing in the mature cells has not been established during the process of ES cell differentiation.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2007年第45期3203-3207,共5页
National Medical Journal of China
基金
国家自然科学基金重点资助项目(30400148)
国家"973"基金(2006CB0F0603)
北京市自然科学基金(7031002)
北京科技新星计划资助项目(H020821400190)
北京市科委科技计划资助项目(H020220010290)
关键词
胚胎干细胞
细胞分化
神经元
节律基因
Embryonic stem cell
Cell differentiation
Neurons
Circadian gene
作者简介
通讯作者：陈彪，E-mail：pbehan@bjsap．org
