Characterizing Semaphorin Signaling In Vivo Using C. elegans

Methods Mol Biol. 2017:1493:485-498. doi: 10.1007/978-1-4939-6448-2_34.

Abstract

A small model animal Caenorhabditis elegans is particularly suitable for genetic analysis, but cell-type-specific biochemistry is a formidable task in this organism. Here we describe techniques utilizing transgenic C. elegans strains expressing epitope-tagged proteins for analyzing biochemical events, such as protein phosphorylation and formation of protein complex, in a small number of a specific group of cells at a defined stage of development. The techniques are useful for elucidating that C. elegans semaphorin-plexin signaling systems regulate epidermal morphogenesis through modulating TOR signaling and its downstream targets.

Keywords: C. elegans; Epitope-tagging; Immunoblotting; Immunoprecipitation; Phosphorylation; Protein complex formation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Caenorhabditis elegans
  • Caenorhabditis elegans Proteins / metabolism*
  • Semaphorins / metabolism*
  • Signal Transduction*

Substances

  • Caenorhabditis elegans Proteins
  • Semaphorins