title: In vivo single-molecule imaging of syntaxin1A reveals polyphosphoinositide- and activity-dependent trapping in presynaptic nanoclusters.
abstract: Syntaxin1A is organized in nanoclusters that are critical for the docking and priming of secretory vesicles from neurosecretory cells. Whether and how these nanoclusters are affected by neurotransmitter release in nerve terminals from a living organism is unknown. Here we imaged photoconvertible syntaxin1A-mEos2 in the motor nerve terminal of Drosophila larvae by single-particle tracking photoactivation localization microscopy. Opto- and thermo-genetic neuronal stimulation increased syntaxin1A-mEos2 mobility, and reduced the size and molecular density of nanoclusters, suggesting an activity-dependent release of syntaxin1A from the confinement of nanoclusters. Syntaxin1A mobility was increased by mutating its polyphosphoinositide-binding site or preventing SNARE complex assembly via co-expression of tetanus toxin light chain. In contrast, syntaxin1A mobility was reduced by preventing SNARE complex disassembly. Our data demonstrate that polyphosphoinositide favours syntaxin1A trapping, and show that SNARE complex disassembly leads to syntaxin1A dissociation from nanoclusters. Lateral diffusion and trapping of syntaxin1A in nanoclusters therefore dynamically regulate neurotransmitter release.
authors.affiliation: Clem Jones Centre for Ageing Dementia Research, Queensland Brain Institute, The University of Queensland, Brisbane, Queensland 4072, Australia.
authors.forename: Adekunle T
authors.initials: AT
authors.lastname: Bademosi
authors.affiliation: VIB Center for the Biology of Disease, 3000 Leuven, Belgium.|KU Leuven Department of Human Genetics, Leuven Institute for Neurodegenerative Disease (LIND), 3000 Leuven, Belgium.
authors.forename: Elsa
authors.initials: E
authors.lastname: Lauwers
authors.affiliation: Queensland Brain Institute, The University of Queensland, Brisbane, Queensland 4072, Australia.
authors.forename: Pranesh
authors.initials: P
authors.lastname: Padmanabhan
authors.affiliation: School of Biomedical Sciences, The University of Queensland, Brisbane, Queensland 4072, Australia.
authors.forename: Lorenzo
authors.initials: L
authors.lastname: Odierna
authors.affiliation: Clem Jones Centre for Ageing Dementia Research, Queensland Brain Institute, The University of Queensland, Brisbane, Queensland 4072, Australia.
authors.forename: Ye Jin
authors.initials: YJ
authors.lastname: Chai
authors.affiliation: Clem Jones Centre for Ageing Dementia Research, Queensland Brain Institute, The University of Queensland, Brisbane, Queensland 4072, Australia.
authors.forename: Andreas
authors.initials: A
authors.lastname: Papadopulos
authors.affiliation: Queensland Brain Institute, The University of Queensland, Brisbane, Queensland 4072, Australia.|School of Mathematics and Physics, The University of Queensland, Brisbane, Queensland 4072, Australia.
authors.forename: Geoffrey J
authors.initials: GJ
authors.lastname: Goodhill
authors.affiliation: VIB Center for the Biology of Disease, 3000 Leuven, Belgium.|KU Leuven Department of Human Genetics, Leuven Institute for Neurodegenerative Disease (LIND), 3000 Leuven, Belgium.
authors.forename: Patrik
authors.initials: P
authors.lastname: Verstreken
authors.affiliation: Queensland Brain Institute, The University of Queensland, Brisbane, Queensland 4072, Australia.
authors.forename: Bruno
authors.initials: B
authors.lastname: van Swinderen
authors.affiliation: Clem Jones Centre for Ageing Dementia Research, Queensland Brain Institute, The University of Queensland, Brisbane, Queensland 4072, Australia.
authors.forename: Frédéric A
authors.initials: FA
authors.lastname: Meunier
mesh_terms: D000818:Animals; D001665:Binding Sites; D004058:Diffusion; D029721:Drosophila Proteins / Q000235:genetics* / Q000378:metabolism; D004331:Drosophila melanogaster / Q000166:cytology / Q000502:physiology*; D005786:Gene Expression Regulation; D007814:Larva / Q000166:cytology / Q000502:physiology; D008666:Metalloendopeptidases / Q000235:genetics / Q000378:metabolism; D057054:Molecular Imaging / Q000379:methods; D009046:Motor Neurons / Q000378:metabolism / Q000648:ultrastructure; D062308:Optogenetics; D018129:Phosphatidylinositol Phosphates / Q000378:metabolism*; D011485:Protein Binding; D021381:Protein Transport; D050600:SNARE Proteins / Q000235:genetics / Q000378:metabolism; D013569:Synapses / Q000378:metabolism / Q000648:ultrastructure; D009435:Synaptic Transmission; D050827:Syntaxin 1 / Q000235:genetics* / Q000378:metabolism; D013744:Tetanus Toxin / Q000235:genetics / Q000378:metabolism
chemical_list: D029721:Drosophila Proteins; D018129:Phosphatidylinositol Phosphates; D050600:SNARE Proteins; D050827:Syntaxin 1; D013744:Tetanus Toxin; D008666:Metalloendopeptidases; C093556:zinc-endopeptidase, tetanus neurotoxin