title: In vivo single-molecule imaging identifies altered dynamics of calcium channels in dystrophin-mutant C. elegans.
abstract: Single-molecule (SM) fluorescence microscopy allows the imaging of biomolecules in cultured cells with a precision of a few nanometres but has yet to be implemented in living adult animals. Here we used split-GFP (green fluorescent protein) fusions and complementation-activated light microscopy (CALM) for subresolution imaging of individual membrane proteins in live Caenorhabditis elegans (C. elegans). In vivo tissue-specific SM tracking of transmembrane CD4 and voltage-dependent Ca(2+) channels (VDCC) was achieved with a precision of 30 nm within neuromuscular synapses and at the surface of muscle cells in normal and dystrophin-mutant worms. Through diffusion analyses, we reveal that dystrophin is involved in modulating the confinement of VDCC within sarcolemmal membrane nanodomains in response to varying tonus of C. elegans body-wall muscles. CALM expands the applications of SM imaging techniques beyond the petri dish and opens the possibility to explore the molecular basis of homeostatic and pathological cellular processes with subresolution precision, directly in live animals.
authors.affiliation: University Claude Bernard Lyon 1, CGphiMC UMR CNRS 5534, Villeurbanne 69622, France.
authors.forename: Hong
authors.initials: H
authors.lastname: Zhan
authors.affiliation: Department of Biological Sciences, Dana and David Dornsife College of Letters, Arts and Sciences, University of Southern California, Los Angeles, California 90089, USA.
authors.forename: Ramunas
authors.initials: R
authors.lastname: Stanciauskas
authors.affiliation: Division of Electron Microscopy, Biocenter of the University of Würzburg, Am Hubland, 97074 Würzburg, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Stigloher
authors.affiliation: Department of Biological Sciences, Dana and David Dornsife College of Letters, Arts and Sciences, University of Southern California, Los Angeles, California 90089, USA.
authors.forename: Kevin
authors.initials: K
authors.lastname: Keomanee-Dizon
authors.affiliation: University Claude Bernard Lyon 1, CGphiMC UMR CNRS 5534, Villeurbanne 69622, France.
authors.forename: Maelle
authors.initials: M
authors.lastname: Jospin
authors.affiliation: University Claude Bernard Lyon 1, CGphiMC UMR CNRS 5534, Villeurbanne 69622, France.
authors.forename: Jean-Louis
authors.initials: JL
authors.lastname: Bessereau
authors.affiliation: 1] Department of Biological Sciences, Dana and David Dornsife College of Letters, Arts and Sciences, University of Southern California, Los Angeles, California 90089, USA [2] Department of Chemistry, Dana and David Dornsife College of Letters, Arts and Sciences, University of Southern California, Los Angeles, California 90089, USA [3] Department of Physics and Astronomy, Dana and David Dornsife College of Letters, Arts and Sciences, University of Southern California, 1050 Childs way, Los Angeles, California 90089, USA.
authors.forename: Fabien
authors.initials: F
authors.lastname: Pinaud
mesh_terms: D000818:Animals; D017173:Caenorhabditis elegans / Q000235:genetics*; D029742:Caenorhabditis elegans Proteins / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism; D015220:Calcium Channels / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D004058:Diffusion; D016189:Dystrophin / Q000235:genetics*; D031541:Fluorescence Resonance Energy Transfer; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006706:Homeostasis; D008565:Membrane Proteins / Q000378:metabolism; D008856:Microscopy, Fluorescence; D009154:Mutation; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012508:Sarcolemma / Q000378:metabolism
chemical_list: D029742:Caenorhabditis elegans Proteins; D015220:Calcium Channels; D016189:Dystrophin; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D002118:Calcium