Evaluating Spindle Assembly Checkpoint Competence in Mouse Oocytes Using Immunoblotting

The spindle assembly checkpoint (SAC) is a quality control mechanism for overseeing the fidelity of chromosome segregation. By modulating the activity of the anaphase-promoting complex or cyclosome (APC/C), the SAC sets the timing of anaphase-onset by co-ordinating the timely destruction of key proteins with the completion of chromosome alignment. How mammalian oocytes regulate chromosome segregation during the first meiotic division (meiosis I) is of immense importance as mis-segregation at this crucial stage in human oocytes underpins the majority of human aneuploidy and birth defects. In recent years, the SAC has been shown to be indispensable for the accuracy of meiosis I chromosome segregation. Here, I describe a technique based on immunoblotting for evaluating SAC competence during meiosis I in mouse oocytes.