Analysis of membrane-bound organelles

Methods Cell Biol. 2012:107:239-63. doi: 10.1016/B978-0-12-394620-1.00008-4.

Abstract

This chapter describes methods for studying membrane traffic and organelle biogenesis in Caenorhabditis elegans. These processes have traditionally been studied with yeast or mammalian cells, but C. elegans is emerging as an attractive alternative model system for cell biologists. C. elegans is well known for the ease of manipulation through classic and molecular genetic techniques. In addition, C. elegans is transparent, so fluorescent proteins can be observed in live animals. These properties have aided the development of functional assays for tracking cell biological processes in situ. Localization results obtained with fluorescent proteins can be validated with immunofluorescence and with biochemical methods, such as subcellular fractionation, adapted from methods developed for other organisms. C. elegans thus combines powerful genetics with a range of cell biological techniques to study subcellular processes in a tractable multicellular organism.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biological Transport
  • Biomarkers / metabolism
  • Caenorhabditis elegans / cytology
  • Caenorhabditis elegans / physiology*
  • Caenorhabditis elegans Proteins / genetics
  • Caenorhabditis elegans Proteins / metabolism
  • Cell Extracts / chemistry
  • Cell Membrane / metabolism
  • Cell Membrane / ultrastructure*
  • Endocytosis
  • Female
  • Fluorescent Antibody Technique / methods
  • Fluorescent Dyes
  • Green Fluorescent Proteins
  • Lysosomes / metabolism
  • Lysosomes / ultrastructure*
  • Microscopy, Fluorescence / methods*
  • Mitochondria / metabolism
  • Mitochondria / ultrastructure*
  • Oocytes / metabolism
  • Oocytes / ultrastructure*
  • Photobleaching

Substances

  • Biomarkers
  • Caenorhabditis elegans Proteins
  • Cell Extracts
  • Fluorescent Dyes
  • Green Fluorescent Proteins