Romé Pierre, Ohkura Hiroyuki
Wellcome Centre for Cell Biology, School of Biological Sciences, University of Edinburgh, Edinburgh, United Kingdom.
Wellcome Centre for Cell Biology, School of Biological Sciences, University of Edinburgh, Edinburgh, United Kingdom.
Methods Cell Biol. 2018;145:237-248. doi: 10.1016/bs.mcb.2018.03.026. Epub 2018 Apr 11.
Studies using Drosophila have played pivotal roles in advancing our understanding of molecular mechanisms of mitosis throughout the past decades, due to the short generation time and advanced genetic research of this organism. Drosophila is also an excellent model to study female meiosis in oocytes. Pathways such as the acentrosomal assembly of the meiotic spindle in oocytes are conserved from fly to humans. Collecting and manipulating large Drosophila oocytes for microscopy and biochemistry are both time and cost efficient, offering advantages over mouse or human oocytes. Therefore, Drosophila oocytes serve as an excellent platform for molecular studies of female meiosis using a combination of genetics, microscopy, and biochemistry. Here we describe key methods to observe the formation of the meiotic spindle either in fixed or in live oocytes. Moreover, biochemical methods are described to identify protein-protein interactions in vivo.
在过去几十年里,由于果蝇的世代周期短且遗传研究先进,利用果蝇进行的研究在推动我们对有丝分裂分子机制的理解方面发挥了关键作用。果蝇也是研究卵母细胞中雌性减数分裂的优秀模型。卵母细胞中减数分裂纺锤体的无中心体组装等途径在从果蝇到人类中都是保守的。收集和操作大量果蝇卵母细胞用于显微镜检查和生物化学研究既节省时间又成本效益高,比小鼠或人类卵母细胞具有优势。因此,果蝇卵母细胞是使用遗传学、显微镜检查和生物化学相结合的方法对雌性减数分裂进行分子研究的优秀平台。在这里,我们描述了在固定或活卵母细胞中观察减数分裂纺锤体形成的关键方法。此外,还描述了用于鉴定体内蛋白质 - 蛋白质相互作用的生物化学方法。