Morphogenetic Analysis of Changing Cell Associations Following Release of 2-cell and 4-cell Mouse Embryos from Cleavage Arrest

Two-cell and four-cell mouse embryos were cultured in Cytochalasin D (CD) for 40-48 h. They were fixed for light and electron microscopy at various times after washing off the CD. Cleavage-arrested embryos in CD had well separated blastomeres but by 1 h from washing the embryos had compacted, in most cases without undergoing cell division. By 2 h after release from arrest one blastomere of the 2-cell arrested embryos had become crescent shaped and at 4-5 h the crescent-shaped blastomere had started to spread over the surface of the other rounded blastomere. This process continued until by 16-24 h from explantation to fresh medium one blastomere had almost completely engulfed the other. A similar process occurred in 4-cell arrested and released embryos. At this stage the embryos had accumulated fluid and become blastocyst-like vesicles. In 20% of 2-cell and 4-cell embryos one or two blastomeres underwent one cell division after release from arrest. Serial sections of these embryos lead to the conclusion that one or both progeny of the first cell to divide tended to be engulfed by the later dividing or non-dividing cell(s). These results are discussed in relation to the differentiation of ICM and trophectoderm in blastocysts.