title: Methods for Studying Movement of Molecules Within Cilia.
abstract: The assembly of cilia and eukaryotic flagella (interchangeable terms) requires the import of numerous proteins from the cell body into the growing organelle. Proteins move into and inside cilia by diffusion and by motor-based intraflagellar transport (IFT). Many aspects of ciliary protein transport such as the distribution of unloading sites and the frequency of transport can be analyzed using direct in vivo imaging of fluorescently tagged proteins. Here, we will describe how to use total internal reflection fluorescence microcopy (TIRFM) to analyze protein transport in the flagella of the unicellular alga Chlamydomonas reinhardtii, a widely used model for cilia and cilia-related disease.
authors: Department of Cellular Biology, University of Georgia, 635 Biological Science Building, 1000 Cedar Street, Athens, GA, 30602, USA. lechtrek@uga.edu.
authors: Karl F
authors: KF
authors: Lechtreck
mesh_terms: D001692:Biological Transport; D002923:Cilia / Q000378:metabolism*; D005407:Flagella / Q000378:metabolism*; D015870:Gene Expression; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics; D008856:Microscopy, Fluorescence; D057054:Molecular Imaging; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012038:Regeneration
chemical_list: D011993:Recombinant Fusion Proteins
keywords: Cilia; Flagella; Green fluorescent protein; Intraflagellar transport (IFT); Total internal reflection fluorescence microscopy (TIRFM)