In vivo imaging of C. elegans endocytosis.
Over the past decade, the early Caenorhabditis elegans embryo has proven to be a useful animal model to study a variety of membrane trafficking events, at least in part due to its large size, optical transparency, and ease of manipulation. Importantly, the stereotypic nature of membrane remodeling that occurs during early embryogenesis has enabled quantitative measurement of endocytic flux. In the absence of exogenous stimulation, resumption of the cell cycle triggered by fertilization is coupled to a dramatic redistribution of plasma membrane content. Numerous proteins are rapidly internalized via clathrin-mediated endocytosis, and the fate of these cargoes can be followed precisely using live imaging in utero. Key to these studies is the maintenance of animal health and their immobilization, which can become technically challenging during extended imaging sessions. Here we highlight recent advances in live imaging techniques that have facilitated the interrogation of endocytic transport in live animals. We focus on the use of transgenic C. elegans strains that stably express fluorescently-tagged proteins, including components of the endosomal system and cargo molecules that traverse this network of membranes. Our findings demonstrate the utility of the C. elegans embryo in defining regulatory mechanisms that control the numerous steps of endocytic trafficking.
Department of Biomolecular Chemistry, University of Wisconsin-Madison School of Medicine and Public Health, Madison, WI 53706, USA.
Lei
L
Wang
Department of Biomolecular Chemistry, University of Wisconsin-Madison School of Medicine and Public Health, Madison, WI 53706, USA. Electronic address: audhya@wisc.edu.
Anjon
A
Audhya
D000818:Animals; D030801:Animals, Genetically Modified; D017173:Caenorhabditis elegans; D002462:Cell Membrane / Q000378:metabolism; D004625:Embryo, Nonmammalian; D004705:Endocytosis; D011992:Endosomes / Q000378:metabolism; D057054:Molecular Imaging / Q000379:methods*; D021381:Protein Transport / Q000235:genetics
Adaptor protein complex; Clathrin; ESCRT; Rab-type GTPase; Recycling