Askjaer Peter, Galy Vincent, Meister Peter
Andalusian Center for Developmental Biology (CABD), CSIC/Junta de Andalucía/Universidad Pablo de Olavide, Carretera de Utrera, Seville, Spain.
Sorbonne Universités, UPMC, Univ Paris 06, UMR7622, IBPS, F-75005 Paris, France; CNRS, UMR7622, IBPS, F-75005 Paris, France.
Methods Cell Biol. 2014;122:277-310. doi: 10.1016/B978-0-12-417160-2.00013-8.
The nematode Caenorhabditis elegans is characterized by many features that make it highly attractive to study nuclear pore complexes (NPCs) and nucleocytoplasmic transport. NPC composition and structure are highly conserved in nematodes and being amenable to a variety of genetic manipulations, key aspects of nuclear envelope dynamics can be observed in great details during breakdown, reassembly, and interphase. In this chapter, we provide an overview of some of the most relevant modern techniques that allow researchers unfamiliar with C. elegans to embark on studies of nucleoporins in an intact organism through its development from zygote to aging adult. We focus on methods relevant to generate loss-of-function phenotypes and their analysis by advanced microscopy. Extensive references to available reagents, such as mutants, transgenic strains, and antibodies are equally useful to scientists with or without prior C. elegans or nucleoporin experience.
线虫秀丽隐杆线虫具有许多特性，使其成为研究核孔复合体（NPC）和核质运输极具吸引力的对象。NPC的组成和结构在线虫中高度保守，并且易于进行各种基因操作，因此在核膜解体、重新组装和间期过程中，可以非常详细地观察到核膜动态的关键方面。在本章中，我们概述了一些最相关的现代技术，这些技术使不熟悉秀丽隐杆线虫的研究人员能够通过从受精卵到衰老成虫的发育过程，在完整生物体中开展核孔蛋白的研究。我们重点介绍与产生功能丧失表型及其通过先进显微镜分析相关的方法。对于有或没有秀丽隐杆线虫或核孔蛋白研究经验的科学家而言，广泛引用可用试剂（如突变体、转基因菌株和抗体）同样有用。
