In vivo single-molecule imaging identifies altered dynamics of calcium channels in dystrophin-mutant C. elegans

Hong Zhan, Ramunas Stanciauskas, Christian Stigloher, Kevin Keomanee-Dizon (), Maelle Jospin, Jean-Louis Bessereau and Fabien Pinaud ()
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Hong Zhan: University Claude Bernard Lyon 1, CGphiMC UMR CNRS 5534
Ramunas Stanciauskas: Dana and David Dornsife College of Letters, Arts and Sciences, University of Southern California
Christian Stigloher: Biocenter of the University of Würzburg
Kevin Keomanee-Dizon: Dana and David Dornsife College of Letters, Arts and Sciences, University of Southern California
Maelle Jospin: University Claude Bernard Lyon 1, CGphiMC UMR CNRS 5534
Jean-Louis Bessereau: University Claude Bernard Lyon 1, CGphiMC UMR CNRS 5534
Fabien Pinaud: Dana and David Dornsife College of Letters, Arts and Sciences, University of Southern California

Nature Communications, 2014, vol. 5, issue 1, 1-12

Abstract: 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 Ca2+ 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.

Date: 2014
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DOI: 10.1038/ncomms5974

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