title: Modern tools to study nuclear pore complexes and nucleocytoplasmic transport in Caenorhabditis elegans.
abstract: 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.
authors.affiliation: Andalusian Center for Developmental Biology (CABD), CSIC/Junta de Andalucía/Universidad Pablo de Olavide, Carretera de Utrera, Seville, Spain.
authors.forename: Peter
authors.initials: P
authors.lastname: Askjaer
authors.affiliation: Sorbonne Universités, UPMC, Univ Paris 06, UMR7622, IBPS, F-75005 Paris, France; CNRS, UMR7622, IBPS, F-75005 Paris, France.
authors.forename: Vincent
authors.initials: V
authors.lastname: Galy
authors.affiliation: Cell fate and Nuclear Organization, Institute of Cell Biology, University of Bern, Baltzerstrasse 4, Bern, Switzerland.
authors.forename: Peter
authors.initials: P
authors.lastname: Meister
mesh_terms: D021581:Active Transport, Cell Nucleus; D000818:Animals; D017173:Caenorhabditis elegans / Q000235:genetics*; D029742:Caenorhabditis elegans Proteins / Q000378:metabolism*; D002843:Chromatin / Q000378:metabolism; D004625:Embryo, Nonmammalian / Q000166:cytology / Q000196:embryology; D005455:Fluorescent Antibody Technique / Q000379:methods; D046529:Microscopy, Electron, Transmission / Q000379:methods; D022022:Nuclear Pore / Q000378:metabolism*; D028861:Nuclear Pore Complex Proteins / Q000032:analysis*; D034622:RNA Interference; D034741:RNA, Small Interfering
chemical_list: D029742:Caenorhabditis elegans Proteins; D002843:Chromatin; D028861:Nuclear Pore Complex Proteins; D034741:RNA, Small Interfering
keywords: CRISPR-Cas9; Caenorhabditis elegans; Live imaging; Nuclear pore complex; Nucleocytoplasmic transport; Nucleoporin; RNAi; Super-resolution microscopy; npc; npp