Laband Kimberley, Lacroix Benjamin, Edwards Frances, Canman Julie C, Dumont Julien
Institut Jacques Monod, CNRS, UMR 7592, University Paris Diderot, Paris, France.
Columbia University Medical Center, New York, NY, United States.
Methods Cell Biol. 2018;145:217-236. doi: 10.1016/bs.mcb.2018.03.025. Epub 2018 Apr 11.
Caenorhabditis elegans is a self-fertilizing hermaphroditic worm. A single C. elegans worm therefore produces both male and female gametes that fuse to generate embryos. While sperm production stops at the end of the C. elegans larval development, oocytes are continuously generated and fertilized during the entire reproductive life of the adult worm. The molecular and cellular mechanisms involved in gametogenesis and the early embryonic divisions are highly conserved between worms and humans; thus C. elegans is a powerful model to study meiotic and mitotic cell divisions in a metazoan system. Additionally, the optical transparency of the worm combined with the ease of the genome-editing methods can be used to easily follow the subcellular behavior of any fluorescently tagged protein of interest using light microscopy approaches. Here we describe two methods for preparing live samples to study oocyte meiotic and early embryonic mitotic divisions by confocal microscopy in C. elegans.
秀丽隐杆线虫是一种自体受精的雌雄同体蠕虫。因此，单个秀丽隐杆线虫蠕虫会产生雄性和雌性配子，它们融合形成胚胎。虽然秀丽隐杆线虫幼虫发育结束时精子产生停止，但在成年蠕虫的整个生殖生命过程中，卵母细胞会持续产生并受精。配子发生和早期胚胎分裂所涉及的分子和细胞机制在蠕虫和人类之间高度保守；因此，秀丽隐杆线虫是研究后生动物系统中减数分裂和有丝分裂细胞分裂的有力模型。此外，蠕虫的光学透明性与基因组编辑方法的简便性相结合，可通过光学显微镜方法轻松追踪任何感兴趣的荧光标记蛋白的亚细胞行为。在这里，我们描述了两种制备活样本的方法，用于通过共聚焦显微镜研究秀丽隐杆线虫的卵母细胞减数分裂和早期胚胎有丝分裂。
