Guangdong Key Laboratory of Male Reproductive Medicine and Genetics, Peking University Shenzhen Hospital, The Institute of Urology, Shenzhen PKU-HKUST Medical Center, Shenzhen 518036, Peoples Republic of China.
Cell Biol Int. 2012 May 1;36(5):491-5. doi: 10.1042/CBI20110423.
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 μM 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, Blimp1, 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.
视黄酸（atRA）已知能诱导 mESCs（小鼠胚胎干细胞）在体外分化为 PGCs（原始生殖细胞）。然而，当 mESCs 衍生的类胚体（EBs）被 atRA 诱导时，尚不清楚哪些与 PGC 分化相关的基因发生变化，以及涉及哪些机制。用 1、2 或 5 μM atRA 处理 mESCs 衍生的 EBs 16 h、2 天或 5 天。分别用实时 PCR 和 Western blot 分析检测与 PGC 分化相关的基因（Lin28、Blimp1、Stra8 和 Mvh）和相应蛋白的相对水平。免疫荧光检测蛋白在 EBs 中的位置和分布。基因的表达特征可分为三类：16 h 内迅速达到峰值，然后下降（Stra8、Lin28）；起始时低，然后增加到 5 天达到峰值（Mvh），相对不变（Blimp1）。atRA 处理 2 天或 5 天的 EBs 中低水平表达 Lin28。Lin28 mRNA 水平的变化不影响 Blimp1 mRNA 水平的变化。Stra8/Lin28 的变化与各自 mRNA 水平的相应变化一致，但 Mvh/Blimp1 的变化与各自 mRNA 水平的相应变化不一致。Blimp1 表达可能独立于 atRA 对 PGC 分化的影响。atRA 可能促进 PGC 分化过程中 Lin28 表达水平低的时期开始。
