4D imaging reveals a shift in chromosome segregation dynamics during mouse pre-implantation development.

Cells of the early developing mammalian embryo frequently mis-segregate chromosomes during cell division, causing daughter cells to inherit an erroneous numbers of chromosomes. Why the embryo is so susceptible to errors is unknown, and the mechanisms that embryos employ to accomplish chromosome segregation are poorly understood. Chromosome segregation is performed by the spindle, a fusiform-shaped microtubule-based transient organelle. Here we present a detailed analysis of 4D fluorescence-confocal data sets of live embryos progressing from the one-cell embryo stage through to blastocyst in vitro, providing some of the first mechanistic insights into chromosome segregation in the mammalian embryo. We show that chromosome segregation occurs as a combined result of poleward chromosome motion (anaphase-A) and spindle elongation (anaphase-B), which occur simultaneously at the time of cell division. Unexpectedly, however, regulation of the two anaphase mechanisms changes significantly between the first and second embryonic mitoses. In one-cell embryos, the velocity of anaphase-A chromosome motion and the velocity and overall extent of anaphase-B spindle elongation are significantly constrained compared with later stages. As a result chromosomes are delivered close to the center of the forming two-cell stage blastomeres at the end of the first mitosis. In subsequent divisions, anaphase-B spindle elongation is faster and more extensive, resulting in the delivery of chromosomes to the distal plasma membrane of the newly forming blastomeres. Metaphase spindle length scales with cell size from the two-cell stage onwards, but is substantially shorter in the first mitosis than in the second mitosis, and the duration of mitosis-1 is substantially greater than subsequent divisions. Thus, there is a striking and unexpected shift in the approach to cell division between the first and second mitotic divisions, which likely reflects adaptations to the unique environment within the developing embryo.