Yamaguchi Shinpei, Kimura Hironobu, Tada Masako, Nakatsuji Norio, Tada Takashi
Laboratory of Stem Cell Engineering, Stem Cell Research Center, Institute for Frontier Medical Sciences, Kyoto University, Shogoin, Sakyo-ku, Japan.
Gene Expr Patterns. 2005 Jun;5(5):639-46. doi: 10.1016/j.modgep.2005.03.001. Epub 2005 Apr 9.
Nanog is a newly identified transcriptional factor bearing a homeodomain and expressed in pluripotential cells of preimplantation and early postimplantation embryos, and embryonic stem (ES) and embryonic germ (EG) cells. Knockout experiments indicate that Nanog functions as a key player in maintaining the pluripotency of stem cells. Importantly, Nanog expression is highly expressed in primordial germ cells (PGCs) of E11.5 and E12.5 mouse embryos. However, its temporal and spatial expression pattern and function in germ cells are largely unknown. To address these issues, whole embryos and cryosections of embryos were immunostained with anti-NANOG and anti-STELLA/PGC7 antibodies. NANOG expression, repressed in colonized PGCs of E7.25-E7.5 embryos, became detectable in migrating PGCs of E7.75-E8.0 embryos. Both male and female PGCs migrating in E9.5 and E10.5 embryos and colonizing the genital ridges of E11.5 and E12.5 embryos were positive for NANOG immunostaining, while the NANOG expression pattern differed between the sexes in the later developmental stage. In female gonadal PGCs of E13.5 and E14.5 embryos, NANOG became undetectable in germ cells positive for the synaptonemal complex-specific protein SCP3, while in male PGCs of E14.5-E16.5 embryos, the number of NANOG-positive germ cells drastically decreased during the mitotic arrest. No germ cells positive for NANOG were detectable in testes and ovaries of adult mice. Thus, in germ cell development, NANOG is expressed in proliferating germ cells, in which nuclear reprogramming is progressing.
Nanog是一种新发现的转录因子,带有一个同源结构域,在植入前和植入后早期胚胎的多能细胞以及胚胎干细胞(ES)和胚胎生殖(EG)细胞中表达。基因敲除实验表明,Nanog在维持干细胞的多能性方面起着关键作用。重要的是,Nanog在E11.5和E12.5小鼠胚胎的原始生殖细胞(PGC)中高度表达。然而,其在生殖细胞中的时空表达模式和功能在很大程度上尚不清楚。为了解决这些问题,用抗NANOG和抗STELLA/PGC7抗体对整个胚胎和胚胎冷冻切片进行免疫染色。NANOG表达在E7.25-E7.5胚胎定殖的PGC中受到抑制,在E7.75-E8.0胚胎迁移的PGC中变得可检测到。在E9.5和E10.5胚胎中迁移并定殖到E11.5和E12.5胚胎生殖嵴的雄性和雌性PGC均对NANOG免疫染色呈阳性,而在后期发育阶段,NANOG的表达模式在两性之间有所不同。在E13.5和E14.5胚胎的雌性性腺PGC中,在联会复合体特异性蛋白SCP3呈阳性的生殖细胞中检测不到NANOG,而在E14.5-E16.5胚胎的雄性PGC中,有丝分裂停滞期间NANOG阳性生殖细胞的数量急剧减少。在成年小鼠的睾丸和卵巢中未检测到NANOG阳性的生殖细胞。因此,在生殖细胞发育过程中,NANOG在增殖的生殖细胞中表达,其中核重编程正在进行。