OCT4 is a fundamental component of the molecular circuitry governing pluripotency in vivo and in vitro. To determine how OCT4 establishes and protects the pluripotent lineage in the embryo, we used comparative single-cell transcriptomics and quantitative immunofluorescence on control and OCT4 null blastocyst inner cell masses at two developmental stages. Surprisingly, activation of most pluripotency-associated transcription factors in the early mouse embryo occurs independently of OCT4, with the exception of the JAK/STAT signaling machinery. Concurrently, OCT4 null inner cell masses ectopically activate a subset of trophectoderm-associated genes. Inspection of metabolic pathways implicates the regulation of rate-limiting glycolytic enzymes by OCT4, consistent with a role in sustaining glycolysis. Furthermore, up-regulation of the lysosomal pathway was specifically detected in OCT4 null embryos. This finding implicates a requirement for OCT4 in the production of normal trophectoderm. Collectively, our findings uncover regulation of cellular metabolism and biophysical properties as mechanisms by which OCT4 instructs pluripotency.

OCT4 是控制体内和体外多能性的分子电路的基本组成部分。为了确定 OCT4 如何在胚胎中建立和保护多能谱系，我们对两个发育阶段的对照和 OCT4 缺失囊胚内细胞团使用了比较单细胞转录组学和定量免疫荧光。令人惊讶的是，除了 JAK/STAT 信号机制外，早期小鼠胚胎中大多数多能性相关转录因子的激活独立于 OCT4 发生。同时，OCT4 无效的内细胞团异位激活滋养外胚层相关基因的子集。对代谢途径的检查表明 OCT4 对限速糖酵解酶的调节，这与维持糖酵解的作用一致。此外，在 OCT4 无效胚胎中特别检测到溶酶体途径的上调。这一发现表明正常滋养外胚层的产生需要 OCT4。总的来说，我们的研究结果揭示了细胞代谢和生物物理特性的调节是 OCT4 指导多能性的机制。