During preimplantation development in the mouse, two phenotypically distinct cell populations appear at the 16-cell stage: nonpolarized inner cells that give rise to the inner cell mass and polarized outer cells that give rise mainly to the trophectoderm. The divergence of these two cell lineages is due to asymmetrical cell divisions during the transition from the 8- to the 16 cell stage which can occur following blastomere polarization. During compaction, at the 8-cell stage, cytoplasmic organelles accumulate in the apical domain, a surface pole of microvilli forms, and blastomeres flatten onto one another. During the division from the 8- to the 16-cell stage, the only asymmetrical structure maintained is the pole of microvilli. At the 16-cell stage, only blastomeres inheriting a large part of this apical structure can reestablish a polarized organization. The mechanisms involved in the formation and stabilization of the apical pole of microvilli are still unknown. Ezrin is an actin-associated protein that has been proposed to play a role in the formation of microvillous structures. This led us to study the expression of ezrin during early development of the mouse embryo. We observed that ezrin mRNA and protein are present in the mouse oocyte and throughout preimplantation embryo development, although the amount of protein present decreases continuously during early development, particularly after the 8-cell stage, at the time of compaction. Two isoforms of ezrin phosphorylated on tyrosine residues are present during all of preimplantation development while a third non-tyrosine-phosphorylated isoform appears at the 8-cell stage and its relative amount increases from the 8-cell stage to the blastocyst stage. Before compaction, ezrin is distributed around the cell cortex. However ezrin becomes restricted to the microvilli of the apical pole after compaction. At later stages, ezrin is found in the microvilli of the apical surface of outer cells. Finally, ezrin remains associated with the microvillous pole during the transition from the 8- to 16-cell stage and is found only in the outer cells after division. Thus, ezrin is the first cytocortical protein described that is totally segregated in outer cells at the 16-cell stage after an asymmetrical division.