Analysis of membrane-bound organelles.
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.
Department of Molecular and Cellular Biology, University of Arizona, Tucson, AZ, USA.
Hanna
H
Fares
Alexander M
AM
van der Bliek
D000818:Animals; D001692:Biological Transport; D015415:Biomarkers / Q000378:metabolism; D017173:Caenorhabditis elegans / Q000166:cytology / Q000502:physiology*; D029742:Caenorhabditis elegans Proteins / Q000235:genetics / Q000378:metabolism; D002457:Cell Extracts / Q000737:chemistry; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure*; D004705:Endocytosis; D005260:Female; D005455:Fluorescent Antibody Technique / Q000379:methods; D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins; D008247:Lysosomes / Q000378:metabolism / Q000648:ultrastructure*; D008856:Microscopy, Fluorescence / Q000379:methods*; D008928:Mitochondria / Q000378:metabolism / Q000648:ultrastructure*; D009865:Oocytes / Q000378:metabolism / Q000648:ultrastructure*; D038761:Photobleaching
D015415:Biomarkers; D029742:Caenorhabditis elegans Proteins; D002457:Cell Extracts; D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins