In vivo analysis of recycling endosomes in Caenorhabditis elegans

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

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.

MeSH terms

  • Animals
  • Caenorhabditis elegans / metabolism*
  • Caenorhabditis elegans / ultrastructure
  • Caenorhabditis elegans Proteins / metabolism
  • Endocytosis
  • Endosomes / metabolism*
  • Endosomes / ultrastructure
  • Green Fluorescent Proteins / metabolism
  • Intestinal Mucosa / metabolism
  • Intestinal Mucosa / ultrastructure
  • Microscopy, Fluorescence
  • Protein Transport

Substances

  • Caenorhabditis elegans Proteins
  • Green Fluorescent Proteins