GSE159433
Membrane tension regulated mechanotransduction drives fate transitions in embryonic stem cells
Changes in cell shape and mechanics frequently accompany cell fate transitions. Yet how cellular  mechanics affects the regulatory pathways controlling cell fate is poorly understood. To probe the  interplay between shape, mechanics and fate, we used embryonic stem (ES) cells, which spread as  they undergo early differentiation. We found that this spreading is regulated by a ß-catenin  mediated decrease in RhoA activity and subsequent decrease in the plasma membrane tension.  Strikingly, preventing the membrane tension decrease resulted in early differentiation defects in  ES cells and gastruloids. We further find that the decrease in membrane tension facilitates  endocytosis of FGF signaling components, which activates ERK signaling and directs exit from  the ES cell state. The early differentiation defects we observed can be rescued by increasing Rab5afacilitated  endocytosis. Thus, we show that a mechanically-triggered increase in endocytosis  regulates early differentiation. Our findings are of fundamental importance for understanding how  cell mechanics regulates biochemical signaling, and therefore cell fate. Overall design: mESC from parental cell line E14 wt and from inducible constitutively active Ezrin (iEZR_CA) were seeded on gelatin-coated plates in 2i+LIF (naive medium) or N2B27 (differentiation medium). The expression of Ezrin was induced ~16hr before exit by the addition of doxycycline to the medium. RNA lysates were collected at 24hr and 48hr of exit from the naive and differentiated cells in both cell lines. 3 replicates per condition were processed.