Receptor-mediated Endocytosis in the<i>Caenorhabditis elegans</i>Oocyte

TLDR

The C. elegans oocyte provides a powerful system for genetic studies of receptor‑mediated endocytosis. The study aims to use YP170::GFP transgenic strains to monitor yolk endocytosis in vivo. YP170::GFP reporter strains were employed to assay endocytic protein functions via RNAi and to screen for rme mutants.

Abstract

The Caenorhabditis elegans oocyte is a highly amenable system for forward and reverse genetic analysis of receptor-mediated endocytosis. We describe the use of transgenic strains expressing a vitellogenin::green fluorescent protein (YP170::GFP) fusion to monitor yolk endocytosis by theC. elegans oocyte in vivo. This YP170::GFP reporter was used to assay the functions of C. eleganspredicted proteins homologous to vertebrate endocytosis factors using RNA-mediated interference. We show that the basic components and pathways of endocytic trafficking are conserved between C. elegans and vertebrates, and that this system can be used to test the endocytic functions of any new gene. We also used the YP170::GFP assay to identify rme(receptor-mediated endocytosis) mutants. We describe a new member of the low-density lipoprotein receptor superfamily, RME-2, identified in our screens for endocytosis defective mutants. We show that RME-2 is the C. elegans yolk receptor.