title: Total internal reflection fluorescence microscopy of intraflagellar transport in Tetrahymena thermophila.
abstract: Live imaging has become a powerful tool in studies of ciliary proteins. Tetrahymena thermophila is an established ciliated model with well-developed genetic and biochemical approaches, but its large size, complex shape, and the large number of short and overlapping cilia, have made live imaging of ciliary proteins challenging. Here we describe a method that combines paralysis of cilia by nickel ions and total internal reflection microscopy for live imaging of fluorescent proteins inside cilia of Tetrahymena. Using this method, we quantitatively documented the intraflagellar transport in Tetrahymena.
authors.affiliation: Department of Cellular Biology, University of Georgia, Athens, GA, USA.
authors.forename: Yu-Yang
authors.initials: YY
authors.lastname: Jiang
authors.affiliation: Department of Cellular Biology, University of Georgia, Athens, GA, USA.
authors.forename: Karl
authors.initials: K
authors.lastname: Lechtreck
authors.affiliation: Department of Cellular Biology, University of Georgia, Athens, GA, USA.
authors.forename: Jacek
authors.initials: J
authors.lastname: Gaertig
mesh_terms: D002923:Cilia / Q000378:metabolism*; D004398:Dyneins / Q000378:metabolism; D016547:Kinesins / Q000378:metabolism; D007734:Kymography / Q000379:methods; D008124:Locomotion / Q000502:physiology*; D008856:Microscopy, Fluorescence / Q000379:methods; D009532:Nickel / Q000494:pharmacology; D021381:Protein Transport / Q000502:physiology; D015800:Protozoan Proteins / Q000378:metabolism*; D016808:Tetrahymena thermophila / Q000502:physiology*
chemical_list: D015800:Protozoan Proteins; D009532:Nickel; D004398:Dyneins; D016547:Kinesins
keywords: Cilia; Ciliate; Dynein; IFT; Kinesin
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