Cell-Size-Independent Spindle Checkpoint Failure Underlies Chromosome Segregation Error in Mouse Embryos

Abstract

Chromosome segregation errors during mammalian preimplantation development cause "mosaic" embryos comprising a mixture of euploid and aneuploid cells, which reduce the potential for a successful pregnancy [1-5], but why these errors are common is unknown. In most cells, chromosome segregation error is averted by the spindle assembly checkpoint (SAC), which prevents anaphase-promoting complex (APC/C) activation and anaphase onset until chromosomes are aligned with kinetochores attached to spindle microtubules [6, 7], but little is known about the SAC's role in the early mammalian embryo. In C. elegans, the SAC is weak in early embryos, and it strengthens during early embryogenesis as a result of progressively lessening cell size [8, 9]. Here, using live imaging, micromanipulation, gene knockdown, and pharmacological approaches, we show that this is not the case in mammalian embryos. Misaligned chromosomes in the early mouse embryo can recruit SAC components to mount a checkpoint signal, but this signal fails to prevent anaphase onset, leading to high levels of chromosome segregation error. We find that failure of the SAC to prolong mitosis is not attributable to cell size. We show that mild chemical inhibition of APC/C can extend mitosis, thereby allowing more time for correct chromosome alignment and reducing segregation errors. SAC-APC/C disconnect thus presents a mechanistic explanation for frequent chromosome segregation errors in early mammalian embryos. Moreover, our data provide proof of principle that modulation of the SAC-APC/C axis can increase the likelihood of error-free chromosome segregation in cultured mammalian embryos.

Keywords: mitosis; mosaic embryo aneuploidy; preimplantation embryo; spindle assembly checkpoint.