Effect of all-trans-retinoic acid on the expression of primordial germ cell differentiation-associated genes in mESC-derived EBs
Effect of all-trans-retinoic acid on the expression of primordial germ cell differentiation-associated genes in mESC-derived EBs
复制标题
全反式视黄酸对 mESC 来源的 EB 中原始生殖细胞分化相关基因表达的影响
DOI：
10.1042/cbi20110423
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发表时间：
2012-05-01
影响因子：
3.9
通讯作者：
Cai, Zhi-Ming
中科院分区：
文献类型：
--
作者：
Guo, Xin;Qi, Zheng-Yu;Cai, Zhi-Ming
已知atRA（全反式视黄酸）在体外诱导mESC（小鼠胚胎干细胞）分化为PGC（原始生殖细胞）。然而，目前尚不清楚在PGC分化相关基因发生了什么变化或什么机制时，涉及来自mESC的EB（胚状体）由atRA诱导。将来自mESC的EB用1、2或5 μ M atRA处理16小时、2天或5天。采用Real-time PCR和Western blot方法分别检测PGC分化相关基因（Lin 28、Blimp 1、Stra 8和Mvh）及其蛋白的相对表达水平。免疫荧光法检测EBs中蛋白的定位和分布。基因的表达特征可分为3类：16 h内迅速达到峰值后下降（Stra 8，Lin 28）、先低后高，5 d内达到峰值（Mvh）和相对无变化（Blimp 1）。Lin 28在atRA处理2天或5天的EB中表达水平较低。Lin 28 mRNA水平的变化不影响Blimp 1 mRNA水平的变化。Stra 8/Lin 28的变化与其各自mRNA水平的相应变化一致，但Mvh/Blimp 1的变化与其各自mRNA水平的相应变化不一致。Blimp 1的表达可能不依赖于atRA对PGC分化的影响。atRA可以促进PGC分化期间Lin 28低水平表达的时期的开始。
atRA (all-trans-retinoic acid) is known to induce the differentiation of mESCs (mouse embryonic stem cells) into PGCs (primordial germ cells) in vitro. However, it is not clear as to what changes occur in PGC differentiation-associated genes or what mechanisms are involved when EBs (embryoid bodies) derived from mESCs are induced by atRA. EBs derived from mESCs were treated with 1, 2 or 5 mu M atRA for 16 h, 2 days or 5 days. Real-time PCR and Western blot analysis were performed to detect the relative levels of PGC differentiation-associated genes (Lin28, Blimp 1, Stra8 and Mvh) and the corresponding proteins respectively. Immunofluorescence was used to detect the protein location and distribution in EBs. The expression characteristics of genes could be divided into three categories: rapidly reached the peak value in 16 h and then decreased (Stra8, Lin28), initially low and then increased to reach the peak value in 5 days (Mvh) and relatively unchanged (Blimp1). A low level of Lin28 was expressed in EBs treated with atRA for 2 days or 5 days. The variation in the level of Lin28 mRNA did not influence the change in the level of Blimp1 mRNA. The changes in Stra8/Lin28 were consistent with the corresponding changes in the levels of their respective mRNAs, but the changes for Mvh/Blimp1 were not consistent with the corresponding changes in the levels of their respective mRNAs. Blimp1 expression may be independent of the effect of atRA on PGC differentiation. atRA may promote the start of a period in which there is a low level of Lin28 expression during PGC differentiation.