Optogenetic manipulation identifies the roles of ERK and AKT dynamics in controlling mouse embryonic stem cell exit from pluripotency.

ERK and AKT signaling control pluripotent cell self-renewal versus differentiation. ERK pathway activity over time (i.e., dynamics) is heterogeneous between individual pluripotent cells, even in response to the same stimuli. To analyze potential functions of ERK and AKT dynamics in controlling mouse embryonic stem cell (ESC) fates, we developed ESC lines and experimental pipelines for the simultaneous long-term manipulation and quantification of ERK or AKT dynamics and cell fates. We show that ERK activity duration or amplitude or the type of ERK dynamics (e.g., transient, sustained, or oscillatory) alone does not influence exit from pluripotency, but the sum of activity over time does. Interestingly, cells retain memory of previous ERK pulses, with duration of memory retention dependent on duration of previous pulse length. FGF receptor/AKT dynamics counteract ERK-induced pluripotency exit. These findings improve our understanding of how cells integrate dynamics from multiple signaling pathways and translate them into cell fate cues. [Display omitted] • Optogenetic control of ERK and AKT activity in mouse embryonic stem cells • Pluripotency exit is due to cumulative ERK activity, not duration, amplitude, or shape • ESCs retain memory of ERK pulses with memory duration dependent on pulse duration • Lower AKT activity increases pluripotency exit Arekatla et al. use optogenetics to control ERK and AKT dynamics in mouse embryonic stem cells. Cumulative ERK activity over time—not duration, amplitude, or types of dynamics—induces pluripotency exit, and lower AKT activation, at constant ERK activity, contributes to higher pluripotency exit. [ABSTRACT FROM AUTHOR]