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Methods Cell Biol. 2015;130:181-98. doi: 10.1016/bs.mcb.2015.04.001. Epub 2015 Jun 11.

In vivo analysis of recycling endosomes in Caenorhabditis elegans.

Author information

1
Department of Medical Genetics, School of Basic Medicine and the Collaborative Innovation Center for Brain Science, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, China; Institute for Brain Research, Huazhong University of Science and Technology, Wuhan, Hubei, China.
2
Department of Molecular Biology and Biochemistry, Rutgers University, Piscataway, NJ, USA.

Abstract

The microscopic nematode Caenorhabditis elegans (C. elegans) serves as an excellent animal model for studying membrane traffic. This is due in part to its highly advanced genetics and genomics, and a transparent body that allows the visualization of fluorescently tagged molecules in the physiologically relevant context of the intact organism. Notably, C. elegans oocytes, coelomocytes, and intestinal epithelia have been established as facile cellular models to explore nonpolarized and polarized cell membrane trafficking mechanisms. In this chapter, we describe in vivo C. elegans assays, utilizing fluorescent dyes or proteins, to examine the molecular mechanisms that control endocytosis and endocytic recycling. Tissue-specific, steady-state imaging and associated quantitative analysis allow the identification and interpretation of subcellular events in the intact animal. To better understand the kinetic characteristics of recycling tubules that mediate membrane protein recycling, we describe recently developed dynamic-imaging assays in intestinal epithelial cells. Such methods bring new clarity to the system, helping to elucidate the functional roles of recycling mediators.

KEYWORDS:

Caenorhabditis elegans; Coelomocytes; Endocytic recycling; Endocytic tracers; Endocytosis; Endosome dynamics; Epithelia; Oocyte; Retrograde recycling; Transmembrane protein

PMID:
26360035
DOI:
10.1016/bs.mcb.2015.04.001
[Indexed for MEDLINE]

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