Kyogoku Hirohisa, Yoshida Shuhei, Kitajima Tomoya S
Laboratory for Chromosome Segregation, RIKEN Center for Biosystems Dynamics Research (BDR), Kobe, Japan.
Laboratory for Chromosome Segregation, RIKEN Center for Biosystems Dynamics Research (BDR), Kobe, Japan.
Methods Cell Biol. 2018;144:459-474. doi: 10.1016/bs.mcb.2018.03.038. Epub 2018 May 7.
Meiotic divisions in females occur in fully grown oocytes that have a large cytoplasmic volume. The intracellular processes that are needed to accomplish meiotic divisions, such as spindle formation, chromosome segregation, and polar body extrusion, are controlled by the concerted actions of nuclear and cytoplasmic factors, which exhibit dynamic quantitative and spatiotemporal changes during meiotic maturation. Thus, distinguishing between meiotic controls that are mediated by cytoplasmic factors and those mediated by nuclear factors helps in the understanding of the mechanisms underlying meiotic divisions. Here, we describe a method to artificially modify the number of nuclei and the volume of the cytoplasm of mouse oocytes through cytoplasmic removal, enucleation, and cell fusion. The oocytes generated by this method are viable and undergo reproducible meiotic divisions exhibiting the effects of altered amounts of cytoplasmic and nuclear factors, which can be analyzed by various assays, such as live imaging microscopy.
雌性减数分裂发生在具有大量细胞质体积的完全成熟的卵母细胞中。完成减数分裂所需的细胞内过程，如纺锤体形成、染色体分离和极体排出，受核因子和细胞质因子的协同作用控制，这些因子在减数分裂成熟过程中表现出动态的数量和时空变化。因此，区分由细胞质因子介导的减数分裂控制和由核因子介导的减数分裂控制有助于理解减数分裂的机制。在这里，我们描述了一种通过细胞质去除、去核和细胞融合来人工改变小鼠卵母细胞核数量和细胞质体积的方法。通过这种方法产生的卵母细胞是有活力的，并经历可重复的减数分裂，表现出细胞质和核因子数量改变的影响，这些影响可以通过各种分析方法进行分析，如实时光学显微镜成像。
