Palmieri S L, Peter W, Hess H, Schöler H R
European Molecular Biology Laboratory, Gene Expression Program, Heidelberg, Germany.
Dev Biol. 1994 Nov;166(1):259-67. doi: 10.1006/dbio.1994.1312.
Oct-4 is the earliest expressed gene known to encode a transcription factor which is developmentally regulated during mammalian embryogenesis. In order to understand the role of Oct-4 in early murine embryogenesis, we carried out an analysis of the temporal and spatial pattern of protein expression. We report the presence of Oct-4 protein in cultured cells and murine embryos as determined by immunohistochemistry using confocal microscopy. Oct-4 protein is present in both embryonic stem cells and embryonal carcinoma cells, but is down-regulated following differentiation of these cells by culture in the absence of leukemia inhibitory factor or in the presence of retinoic acid, respectively. In embryos, the protein is found at low levels in unfertilized eggs and is localized predominantly to the pro-nuclei upon their formation following fertilization. The protein is present in the nuclei in all cleavage stages, but following differentiation of the trophectoderm at the blastocyst stage, Oct-4 protein is only expressed in the inner cell mass. The pattern of protein expression to this stage correlates well with previously reported in situ hybridization results; however, a striking difference was seen in primitive endoderm cells which had begun to differentiate and migrate along the inner surface of the trophectoderm. In direct contrast to RNA localization studies which demonstrate that there are only low levels of Oct-4 transcripts in primitive endoderm cells, protein expression in these early migrating cells is higher than in the inner cell mass.
Oct-4是已知最早表达的基因,其编码一种在哺乳动物胚胎发生过程中受发育调控的转录因子。为了了解Oct-4在小鼠早期胚胎发生中的作用,我们对蛋白质表达的时空模式进行了分析。我们报告了通过使用共聚焦显微镜的免疫组织化学测定,在培养细胞和小鼠胚胎中存在Oct-4蛋白。Oct-4蛋白存在于胚胎干细胞和胚胎癌细胞中,但在分别通过在无白血病抑制因子的情况下培养或在视黄酸存在的情况下培养使这些细胞分化后,其表达下调。在胚胎中,该蛋白在未受精卵中含量较低,在受精后原核形成时主要定位于原核。在所有卵裂阶段,该蛋白都存在于细胞核中,但在囊胚期滋养外胚层分化后,Oct-4蛋白仅在内细胞团中表达。到这个阶段的蛋白质表达模式与先前报道的原位杂交结果密切相关;然而,在已经开始分化并沿着滋养外胚层内表面迁移的原始内胚层细胞中观察到了显著差异。与RNA定位研究直接相反,RNA定位研究表明原始内胚层细胞中只有低水平的Oct-4转录本,这些早期迁移细胞中的蛋白质表达高于内细胞团中的表达。