The Two Active X Chromosomes in Female ESCs Block Exit from the Pluripotent State by Modulating the ESC Signaling Network

During early development of female mouse embryos, both X chromosomes are transiently active. X gene dosage is then equalized between the sexes through the process of X chromosome inactivation (XCI). Whether the double dose of X-linked genes in females compared with males leads to sex-specific developmental differences has remained unclear. Using embryonic stem cells with distinct sex chromosome compositions as a model system, we show that two X chromosomes stabilize the naive pluripotent state by inhibiting MAPK and Gsk3 signaling and stimulating the Akt pathway. Since MAPK signaling is required to exit the pluripotent state, differentiation is paused in female cells as long as both X chromosomes are active. By preventing XCI or triggering it precociously, we demonstrate that this differentiation block is released once XX cells have undergone X inactivation. We propose that double X dosage interferes with differentiation, thus ensuring a tight coupling between X chromosome dosage compensation and development. [Display omitted] •The presence of two active X chromosomes delays differentiation of mouse ESCs•Double X dosage inhibits the differentiation-promoting MAPK signaling pathway•Delayed differentiation is associated with reduced de novo DNA methylation•Female ESCs must undergo X inactivation to release the differentiation block Schulz et al. show that before X inactivation occurs in female mouse embryonic stem cells, the presence of two active X chromosomes prevents differentiation via a MAPK signaling pathway.