Mathieu J, Huynh J-R
Institut Curie, Paris, France; CNRS UMR3215, Inserm U934, Paris, France.
Methods Cell Biol. 2017;137:105-118. doi: 10.1016/bs.mcb.2016.03.033. Epub 2016 May 2.
In most species, cytokinesis is blocked in germ cells during at least some stage of their development. Abscission is difficult to assess directly in germ cells which are located in internal organs. Here, we described several indirect and direct methods to monitor the completion of abscission in Drosophila germ line cells. These methods are based on the observation that cells still connected by some cytoplasm share some degree of synchronization of their cell cycle. This synchrony can be detected on fixed tissue (Section 1.1), including using EdU incorporation to label S-phase (Section 1.2). Mitotic synchrony can also be observed using short-term live imaging (Section 1.3). Finally, we describe how the completion of abscission can be monitored using photoactivatable markers diffusing or not between two cells (Section 1.4).
在大多数物种中，胞质分裂在生殖细胞发育的至少某些阶段会受到阻碍。由于生殖细胞位于内部器官中，因此难以直接评估其切割过程。在这里，我们描述了几种间接和直接的方法来监测果蝇生殖系细胞中切割的完成情况。这些方法基于这样的观察结果：仍通过一些细胞质相连的细胞在一定程度上共享其细胞周期的同步性。这种同步性可以在固定组织上检测到（第1.1节），包括使用EdU掺入来标记S期（第1.2节）。也可以使用短期实时成像观察有丝分裂同步性（第1.3节）。最后，我们描述了如何使用可光激活的标记物在两个细胞之间扩散或不扩散来监测切割的完成情况（第1.4节）。
