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

Please always quote using this URN: urn:nbn:de:bvb:20-opus-121125
  • 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 nmSingle-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.show moreshow less

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Metadaten
Author: Hong Zhan, Ramunas Stanciauskas, Christian Stigloher, Kevin K. Dizon, Maelle Jospin, Jean-Luis Bessereau, Fabien Pinaud
URN:urn:nbn:de:bvb:20-opus-121125
Document Type:Journal article
Faculties:Medizinische Fakultät / Theodor-Boveri-Institut für Biowissenschaften
Language:English
Parent Title (English):Nature Communications
Year of Completion:2014
Volume:5
Issue:4974
Source:Nature Communications 5:4974 doi: 10.1038/ncomms5974 (2014).
DOI:https://doi.org/10.1038/ncomms5974
Pubmed Id:http://www.ncbi.nlm.nih.gov/pubmed?term=25232639
Dewey Decimal Classification:6 Technik, Medizin, angewandte Wissenschaften / 61 Medizin und Gesundheit / 610 Medizin und Gesundheit
Release Date:2016/02/17
Licence (German):License LogoCC BY: Creative-Commons-Lizenz: Namensnennung