时钟基因在胚胎干细胞分化为神经元过程中的表达特征  

Expression profile of clock genes during the process of embryonic stem cells differentiation

作　　者：谢淑[1,2,3] 蔡彦宁[1,2,3] 关云谦[2,1] 武文琪[4] 徐艳玲[5,2] 张愚[5,2] 陈彪[2,1,3] 

机构地区：[1]首都医科大学宣武医院神经生物学室,北京100053 [2]教育部重点实验室 [3]国家人类基因组北方研究中心 [4]首都医科大学医学实验与测试中心 [5]首都医科大学宣武医院细胞治疗中心,北京100053

出　　处：《中华医学杂志》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)

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.

关 键 词：胚胎干细胞 细胞分化 神经元 节律基因 

分 类 号：Q344[生物学—遗传学]