Diffusion Dynamics of Glycine Receptors Revealed by Single-Quantum Dot Tracking

Dahan, Maxime; Lévi, Sabine; Luccardini, Camilla; Rostaing, Philippe; Riveau, Béatrice; Triller, Antoine

doi:10.1126/science.1088525

Cited by 1,404 publications

(1,251 citation statements)

References 22 publications

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“…Note the extensive central (C-)-domain (yellow), which is exclusively present in gephyrin trimeric gephyrin may generate a variable postsynaptic scaVold for GlyR recruitment and anchoring, which allows dynamic movement in and out of postsynaptic GlyR clusters and thereby generating plasticity of the postsynaptic response. This conception is in agreement with data from single particle tracking and of quantum dot-labeled GlyRs in the neuronal plasma membrane (Dahan et al 2003;Meier et al 2001). The high degree of sequence-and structural conservation between gephyrin G-and E-domains and the corresponding bacterial, respectively plant enzymes may reXect requirements common for both, moco biosynthesis and inhibitory neurotransmitter cluster formation.…”

Section: Analysis Of Gephyrin Dewcient Micesupporting

confidence: 85%

“…Gephyrin aggregates would subsequently trap GlyRs diVusing in the plane of the plasma membrane by decreasing their lateral mobility. This prediction was impressively conWrmed by Meier et al (2001) and very elegantly reWned by Dahan et al (2003), who studied individual respectively quantum dotconjugated GlyRs to follow their movements on the neuronal cell surface in real time. In membrane areas devoid of gephyrin clusters, GlyRs were mostly freely diVusing.…”

Section: Synapse Formation and Plasticitymentioning

confidence: 74%

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Molecular architecture of glycinergic synapses

Dresbach

Nawrotzki

Kremer

et al. 2008

Histochem Cell Biol

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Synapses can be considered chemical machines, which are optimized for fast and repeated exocytosis of neurotransmitters from presynaptic nerve terminals and the reliable electrical or chemical transduction of neurotransmitter binding to the appropriate receptors in the postsynaptic membrane. Therefore, synapses share a common repertoire of proteins like, e.g., the release machinery and certain cell adhesion molecules. This basic repertoire must be extended in order to generate speciWcity of neurotransmission and allow plastic changes, which are considered the basis of developmental and/or learning processes. Here, we focus on these complementary molecules located in the presynaptic terminal and postsynaptic membrane specializations of glycinergic synapses. Moreover, as speciWcity of neurotransmission in this system is established by the speciWc binding of the neurotransmitter to its receptor, we review the molecular properties of glycine receptor subunits and their assembly into functional glycine receptors with diVerent functional characteristics. The past years have revealed that the molecular machinery underlying inhibitory and especially glycinergic postsynaptic membrane specializations is more complex and dynamic than previously anticipated from morphological studies. The emerging features include structural components as well as signaling modules, which could confer the plasticity required for the proper function of distinct motor and sensory functions.

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Section: Analysis Of Gephyrin Dewcient Micesupporting

confidence: 85%

Section: Synapse Formation and Plasticitymentioning

confidence: 74%

Molecular architecture of glycinergic synapses

Dresbach

Nawrotzki

Kremer

et al. 2008

Histochem Cell Biol

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“…EGFRs tagged with a 40 nm gold particle exhibit a diffusivity (~3 Â 10 À3 mm 2 /s) lower than that of EGFRs tagged with a commercial quantum dot (~0.04 mm 2 /s) (50) or fused with an EGFP (~0.2 mm 2 /s) (106). Although large nanoparticle labels have also been used for high-resolution tracking of a wide range of membrane proteins (such as glycine receptors (107), GPI anchor proteins (17), a-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid and N-methyl-D-aspartate receptors (108), integrins (109), and cystic fibrosis transmembrane conductance regulator channel proteins (110,111)), smaller particle labels (diameter < 10 nm) are recently developed and tested for SPT in a confined space (such as synaptic receptors in synaptic cleft (27)), providing less steric hindrance and reduced cross-linking. To completely eliminate cross-linking, we are currently developing monovalent EGFR labeling techniques based on site-specific biotinylation of membrane receptors (113), monovalent streptavidin (114,115), and monovalent nanoparticles (116).…”

Section: Trade-offs In Temporal Resolution and Probe Sizementioning

confidence: 99%

Segmentation of 3D Trajectories Acquired by TSUNAMI Microscope: An Application to EGFR Trafficking

Liu

Perillo

Liu

et al. 2016

Biophysical Journal

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Whereas important discoveries made by single-particle tracking have changed our view of the plasma membrane organization and motor protein dynamics in the past three decades, experimental studies of intracellular processes using singleparticle tracking are rather scarce because of the lack of three-dimensional (3D) tracking capacity. In this study we use a newly developed 3D single-particle tracking method termed TSUNAMI (Tracking of Single particles Using Nonlinear And Multiplexed Illumination) to investigate epidermal growth factor receptor (EGFR) trafficking dynamics in live cells at 16/43 nm (xy/z) spatial resolution, with track duration ranging from 2 to 10 min and vertical tracking depth up to tens of microns. To analyze the long 3D trajectories generated by the TSUNAMI microscope, we developed a trajectory analysis algorithm, which reaches 81% segment classification accuracy in control experiments (termed simulated movement experiments). When analyzing 95 EGF-stimulated EGFR trajectories acquired in live skin cancer cells, we find that these trajectories can be separated into three groups-immobilization (24.2%), membrane diffusion only (51.6%), and transport from membrane to cytoplasm (24.2%). When EGFRs are membrane-bound, they show an interchange of Brownian diffusion and confined diffusion. When EGFRs are internalized, transitions from confined diffusion to directed diffusion and from directed diffusion back to confined diffusion are clearly seen. This observation agrees well with the model of clathrin-mediated endocytosis.

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“…They obtained ∼5 nm localization within 75 ms integration. 8 Organic dyes have not been bright enough to produce the amount of photons required to localize the center within one nanometer precision. They also photobleach on the order of a second, which is generally not long enough for biological applications.…”

Section: Fionamentioning

confidence: 99%

Fluorescence Imaging with One Nanometer Accuracy: Application to Molecular Motors

2005

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We introduce the technique of FIONA, fluorescence imaging with one nanometer accuracy. This is a fluorescence technique that is able to localize the position of a single dye within ∼1 nm in the x-y plane. It is done simply by taking the point spread function of a single fluorophore excited with wide field illumination and locating the center of the fluorescent spot by a two-dimensional Gaussian fit. We motivate the development of FIONA by unraveling the walking mechanism of the molecular motors myosin V, myosin VI, and kinesin. We find that they all walk in a hand-over-hand fashion.

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Diffusion Dynamics of Glycine Receptors Revealed by Single-Quantum Dot Tracking

Cited by 1,404 publications

References 22 publications

Molecular architecture of glycinergic synapses

Molecular architecture of glycinergic synapses

Segmentation of 3D Trajectories Acquired by TSUNAMI Microscope: An Application to EGFR Trafficking

Fluorescence Imaging with One Nanometer Accuracy: Application to Molecular Motors

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