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In vivo single-molecule imaging identifies altered dynamics of calcium channels in dystrophin-mutant C. elegans
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In vivo single-molecule imaging identifies altered dynamics of calcium channels in dystrophin-mutant C. elegans
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In vivo single-molecule imaging identifies altered dynamics of calcium channels in dystrophin-mutant C. elegans
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Journal Article
In vivo single-molecule imaging identifies altered dynamics of calcium channels in dystrophin-mutant C. elegans
2014
Overview
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.
Single molecule fluorescence microscopy is a powerful technique to study protein dynamics in cells, but it has not been applied to adult animals. The authors use complementation-activated light microscopy in
C. elegans
to discover that dystrophin regulates the diffusion properties of voltage-dependent calcium ion channels at the surface of body-wall muscle cells.
Publisher
Nature Publishing Group UK,Nature Publishing Group
Subject
/ 14
/ 14/35
/ 64/11
/ Animals
/ Caenorhabditis elegans - genetics
/ Caenorhabditis elegans Proteins - metabolism
/ Calcium channels (voltage-gated)
/ Calcium Channels - metabolism
/ Fluorescence Resonance Energy Transfer
/ Green Fluorescent Proteins - metabolism
/ Humanities and Social Sciences
/ Labeling
/ Membrane Proteins - metabolism
/ Muscles
/ Mutants
/ Mutation
/ Peptides
/ Proteins
/ Recombinant Fusion Proteins - metabolism
/ Science
/ Synapses
/ Voltage
