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; Fabien Pinaud

doi:10.1038/ncomms5974

journal article Open Access Sep 18, 2014

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 Fabien Pinaud

Nature Communications Vol. 5 No. 1 · Springer Science and Business Media LLC

View at Publisher Save 10.1038/ncomms5974

Abstract

AbstractSingle-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.

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Published: Sep 18, 2014
Vol/Issue: 5(1)
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Authors

H

Hong Zhan

From Great Ormond Street Hospital for Children NHS Trust (R.C., G.O., T.B., J.C., S.A., R.T., K.G., D.O., A.V., W.Q.) and the UCL Great Ormond Street Institute of Child Health (C.G., F.S., H.Z., A.E., S.A.G., R.P., A.K., W.Q.) — both in London.

Cite This Article

Hong Zhan, Ramunas Stanciauskas, Christian Stigloher, et al. (2014). In vivo single-molecule imaging identifies altered dynamics of calcium channels in dystrophin-mutant C. elegans. Nature Communications, 5(1). https://doi.org/10.1038/ncomms5974

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

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