title: Effect of all-trans-retinoic acid on the expression of primordial germ cell differentiation-associated genes in mESC-derived EBs.
abstract: 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.
authors.affiliation: 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.
authors.forename: Xin
authors.initials: X
authors.lastname: Guo
authors.forename: Zheng-Yu
authors.initials: ZY
authors.lastname: Qi
authors.forename: Yan-Min
authors.initials: YM
authors.lastname: Zhang
authors.forename: Jie
authors.initials: J
authors.lastname: Qin
authors.forename: Guang-Hui
authors.initials: GH
authors.lastname: Cui
authors.forename: Yao-Ting
authors.initials: YT
authors.lastname: Gui
authors.forename: Zhi-Ming
authors.initials: ZM
authors.lastname: Cai
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D002454:Cell Differentiation / Q000235:genetics*; D002478:Cells, Cultured; D053487:DEAD-box RNA Helicases / Q000235:genetics / Q000378:metabolism; D058732:Embryoid Bodies / Q000187:drug effects / Q000378:metabolism*; D053595:Embryonic Stem Cells / Q000166:cytology*; D018507:Gene Expression Regulation, Developmental; D005854:Germ Cells / Q000166:cytology* / Q000187:drug effects; D051379:Mice; D000074462:Positive Regulatory Domain I-Binding Factor 1; D011506:Proteins / Q000235:genetics / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014212:Tretinoin / Q000494:pharmacology*
chemical_list: D048868:Adaptor Proteins, Signal Transducing; C488352:Lin-28 protein, mouse; C513258:Prdm1 protein, mouse; D011506:Proteins; D016601:RNA-Binding Proteins; C102019:Stra8 protein, mouse; D014157:Transcription Factors; D014212:Tretinoin; D000074462:Positive Regulatory Domain I-Binding Factor 1; C090864:Ddx4 protein, mouse; D053487:DEAD-box RNA Helicases