Both Nodal signalling and stochasticity select for prospective distal visceral endoderm in mouse embryos

Anterior-posterior (A-P) polarity of mouse embryos is established by distal visceral endoderm (DVE) at embryonic day (E) 5.5. Lefty1 is expressed first at E3.5 in a subset of epiblast progenitor cells (L1^epi cells) and then in a subset of primitive endoderm cells (L1^dve cells) fated to become DVE. Here we studied how prospective DVE cells are selected. Lefty1 expression in L1^epi and L1^dve cells depends on Nodal signaling. A cell that experiences the highest level of Nodal signaling begins to express Lefty1 and becomes an L1^epi cell. Deletion of Lefty1 alone or together with Lefty2 increased the number of prospective DVE cells. Ablation of L1^epi or L1^dve cells triggered Lefty1 expression in a subset of remaining cells. Our results suggest that selection of prospective DVE cells is both random and regulated, and that a fixed prepattern for the A-P axis does not exist before the blastocyst stage.