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  1. Article ; Online: Cellular and structural insight into dynamin function during endocytic vesicle formation: a tale of 50 years of investigation.

    Perrais, David

    Bioscience reports

    2022  Volume 42, Issue 11

    Abstract: Dynamin is one of the major proteins involved in endocytosis. First identified 50 years ago in a genetic screen in Drosophila melanogaster, it has become a central player in many forms of endocytosis, such as clathrin-mediated endocytosis or synaptic ... ...

    Abstract Dynamin is one of the major proteins involved in endocytosis. First identified 50 years ago in a genetic screen in Drosophila melanogaster, it has become a central player in many forms of endocytosis, such as clathrin-mediated endocytosis or synaptic vesicle endocytosis, as well as other important cellular processes such as actin remodelling. Decades of work using biochemical and structural studies, cell-free assays, live cell imaging, acute inhibition and genetic studies have led to important insights on its mode of action. Dynamin is a remarkable mechano-GTPase, which can do a lot to membranes on its own but which is, in cells, at the centre of a vast protein and lipid network and cannot work in isolation. This review summarizes the main features of dynamin structure and function and its central role in membrane remodelling events, and give an update on the latest results.
    MeSH term(s) Animals ; Clathrin/metabolism ; Drosophila melanogaster/metabolism ; Dynamins/genetics ; Dynamins/metabolism ; Endocytosis/physiology ; Transport Vesicles/metabolism
    Chemical Substances Clathrin ; Dynamins (EC 3.6.5.5)
    Language English
    Publishing date 2022-09-01
    Publishing country England
    Document type Review ; Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 764946-0
    ISSN 1573-4935 ; 0144-8463
    ISSN (online) 1573-4935
    ISSN 0144-8463
    DOI 10.1042/BSR20211227
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article: Imaging of post-synaptic membrane trafficking in neuronal dendrites: progress, limitations, and new developments.

    Perrais, David / Sposini, Silvia / Angibaud, Julie

    Neurophotonics

    2023  Volume 10, Issue 4, Page(s) 44404

    Abstract: Membrane trafficking of post-synaptic cargo is a key determinant of synaptic transmission and synaptic plasticity. We describe here the latest developments in visualizing individual exocytosis and endocytosis events in neurons using pH-sensitive tags. We ...

    Abstract Membrane trafficking of post-synaptic cargo is a key determinant of synaptic transmission and synaptic plasticity. We describe here the latest developments in visualizing individual exocytosis and endocytosis events in neurons using pH-sensitive tags. We show how these tools help decipher the spatial and temporal regulation of membrane trafficking steps during synaptic plasticity.
    Language English
    Publishing date 2023-08-04
    Publishing country United States
    Document type Journal Article
    ZDB-ID 2781943-7
    ISSN 2329-4248 ; 2329-423X
    ISSN (online) 2329-4248
    ISSN 2329-423X
    DOI 10.1117/1.NPh.10.4.044404
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  3. Article ; Online: Endocytose dans les dendrites - Un outil local de régulation de la transmission synaptique.

    Perrais, David / Rosendale, Morgane

    Medecine sciences : M/S

    2017  Volume 33, Issue 11, Page(s) 942–945

    Title translation Endocytosis in dendrites: a local tool to regulate synaptic transmission.
    MeSH term(s) Animals ; Dendrites/physiology ; Endocytosis/physiology ; Homeostasis ; Post-Synaptic Density ; Synapses/physiology ; Synaptic Transmission/physiology
    Language French
    Publishing date 2017-12-04
    Publishing country France
    Document type News
    ZDB-ID 632733-3
    ISSN 1958-5381 ; 0767-0974
    ISSN (online) 1958-5381
    ISSN 0767-0974
    DOI 10.1051/medsci/20173311009
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  4. Article ; Online: Imaging in focus: Imaging the dynamics of endocytosis.

    Rosendale, Morgane / Perrais, David

    The international journal of biochemistry & cell biology

    2017  Volume 93, Page(s) 41–45

    Abstract: Endocytosis, the formation of membrane vesicles from the plasma membrane, is an essential feature of eukaryotic cell biology. Intense research effort has been dedicated to developing methods that can detect endocytosis events with the highest resolution. ...

    Abstract Endocytosis, the formation of membrane vesicles from the plasma membrane, is an essential feature of eukaryotic cell biology. Intense research effort has been dedicated to developing methods that can detect endocytosis events with the highest resolution. We have classified these methods into four families. They exploit the physical properties of endocytosis, namely: 1. Distinguishing extracellular from internalised cargo in fixed samples, 2. Monitoring endosomal acidification, 3. Measuring the turnover of endocytic zones and 4. Detecting vesicle scission. The last three families, all based on fluorescence imaging, are used to study endocytosis in living cells. We discuss the advantages and limitations of these methods and conclude on the future developments required to tackle the upcoming challenges in this fundamental field of cell biology.
    Language English
    Publishing date 2017-10-19
    Publishing country Netherlands
    Document type Journal Article
    ZDB-ID 1228429-4
    ISSN 1878-5875 ; 1357-2725
    ISSN (online) 1878-5875
    ISSN 1357-2725
    DOI 10.1016/j.biocel.2017.10.010
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  5. Article ; Online: Synaptotagmin 1-triggered lipid signaling facilitates coupling of exo- and endocytosis.

    Bolz, Svenja / Kaempf, Natalie / Puchkov, Dmytro / Krauss, Michael / Russo, Giulia / Soykan, Tolga / Schmied, Christopher / Lehmann, Martin / Müller, Rainer / Schultz, Carsten / Perrais, David / Maritzen, Tanja / Haucke, Volker

    Neuron

    2023  Volume 111, Issue 23, Page(s) 3900

    Language English
    Publishing date 2023-12-06
    Publishing country United States
    Document type Journal Article ; Published Erratum
    ZDB-ID 808167-0
    ISSN 1097-4199 ; 0896-6273
    ISSN (online) 1097-4199
    ISSN 0896-6273
    DOI 10.1016/j.neuron.2023.11.003
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  6. Article ; Online: Synaptotagmin 1-triggered lipid signaling facilitates coupling of exo- and endocytosis.

    Bolz, Svenja / Kaempf, Natalie / Puchkov, Dmytro / Krauss, Michael / Russo, Giulia / Soykan, Tolga / Schmied, Christopher / Lehmann, Martin / Müller, Rainer / Schultz, Carsten / Perrais, David / Maritzen, Tanja / Haucke, Volker

    Neuron

    2023  Volume 111, Issue 23, Page(s) 3765–3774.e7

    Abstract: Exocytosis and endocytosis are essential physiological processes and are of prime importance for brain function. Neurotransmission depends on the ... ...

    Abstract Exocytosis and endocytosis are essential physiological processes and are of prime importance for brain function. Neurotransmission depends on the Ca
    MeSH term(s) Animals ; Mice ; Endocytosis/physiology ; Exocytosis/physiology ; Lipids ; Synaptic Transmission ; Synaptic Vesicles/metabolism ; Synaptotagmin I/genetics ; Synaptotagmin I/metabolism
    Chemical Substances Lipids ; Synaptotagmin I ; Syt1 protein, mouse
    Language English
    Publishing date 2023-09-21
    Publishing country United States
    Document type Journal Article
    ZDB-ID 808167-0
    ISSN 1097-4199 ; 0896-6273
    ISSN (online) 1097-4199
    ISSN 0896-6273
    DOI 10.1016/j.neuron.2023.08.016
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  7. Article ; Online: Rational Engineering of an Improved Genetically Encoded pH Sensor Based on Superecliptic pHluorin.

    Shen, Yi / Wen, Yurong / Sposini, Silvia / Vishwanath, Anjali Amrapali / Abdelfattah, Ahmed S / Schreiter, Eric R / Lemieux, M Joanne / de Juan-Sanz, Jaime / Perrais, David / Campbell, Robert E

    ACS sensors

    2023  Volume 8, Issue 8, Page(s) 3014–3022

    Abstract: Genetically encoded pH sensors based on fluorescent proteins are valuable tools for the imaging of cellular events that are associated with pH changes, such as exocytosis and endocytosis. Superecliptic pHluorin (SEP) is a pH-sensitive green fluorescent ... ...

    Abstract Genetically encoded pH sensors based on fluorescent proteins are valuable tools for the imaging of cellular events that are associated with pH changes, such as exocytosis and endocytosis. Superecliptic pHluorin (SEP) is a pH-sensitive green fluorescent protein (GFP) variant widely used for such applications. Here, we report the rational design, development, structure, and applications of Lime, an improved SEP variant with higher fluorescence brightness and greater pH sensitivity. The X-ray crystal structure of Lime supports the mechanistic rationale that guided the introduction of beneficial mutations. Lime provides substantial improvements relative to SEP for imaging of endocytosis and exocytosis. Furthermore, Lime and its variants are advantageous for a broader range of applications including the detection of synaptic release and neuronal voltage changes.
    MeSH term(s) Green Fluorescent Proteins/chemistry ; Neurons/metabolism ; Hydrogen-Ion Concentration
    Chemical Substances PHluorin ; Green Fluorescent Proteins (147336-22-9) ; lime (C7X2M0VVNH)
    Language English
    Publishing date 2023-07-23
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ISSN 2379-3694
    ISSN (online) 2379-3694
    DOI 10.1021/acssensors.3c00484
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  8. Article ; Online: Imaging endocytic vesicle formation at high spatial and temporal resolutions with the pulsed-pH protocol.

    Sposini, Silvia / Rosendale, Morgane / Claverie, Léa / Van, Thi Nhu Ngoc / Jullié, Damien / Perrais, David

    Nature protocols

    2020  Volume 15, Issue 9, Page(s) 3088–3104

    Abstract: Endocytosis is a fundamental process occurring in all eukaryotic cells. Live cell imaging of endocytosis has helped to decipher many of its mechanisms and regulations. With the pulsed-pH (ppH) protocol, one can detect the formation of individual ... ...

    Abstract Endocytosis is a fundamental process occurring in all eukaryotic cells. Live cell imaging of endocytosis has helped to decipher many of its mechanisms and regulations. With the pulsed-pH (ppH) protocol, one can detect the formation of individual endocytic vesicles (EVs) with an unmatched temporal resolution of 2 s. The ppH protocol makes use of cargo protein (e.g., the transferrin receptor) coupled to a pH-sensitive fluorescent protein, such as superecliptic pHluorin (SEP), which is brightly fluorescent at pH 7.4 but not fluorescent at pH <6.0. If the SEP moiety is at the surface, its fluorescence will decrease when cells are exposed to a low pH (5.5) buffer. If the SEP moiety has been internalized, SEP will remain fluorescent even during application of the low pH buffer. Fast perfusion enables the complete exchange of low and high pH extracellular solutions every 2 s, defining the temporal resolution of the technique. Unlike other imaging-based endocytosis assays, the ppH protocol detects EVs without a priori hypotheses on the dynamics of vesicle formation. Here, we explain how the ppH protocol quantifies the endocytic activity of living cells and the recruitment of associated proteins in real time. We provide a step-by-step procedure for expression of the reporter proteins with transient transfection, live cell image acquisition with synchronized pH changes and automated analysis. The whole protocol can be performed in 2 d to provide quantitative information on the endocytic process being studied.
    MeSH term(s) Animals ; Cell Line ; Humans ; Hydrogen-Ion Concentration ; Mice ; Molecular Imaging/methods ; Spatio-Temporal Analysis ; Transport Vesicles/metabolism
    Keywords covid19
    Language English
    Publishing date 2020-08-17
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2244966-8
    ISSN 1750-2799 ; 1754-2189
    ISSN (online) 1750-2799
    ISSN 1754-2189
    DOI 10.1038/s41596-020-0371-z
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  9. Article ; Online: Spatial and Temporal Regulation of Receptor Endocytosis in Neuronal Dendrites Revealed by Imaging of Single Vesicle Formation.

    Rosendale, Morgane / Jullié, Damien / Choquet, Daniel / Perrais, David

    Cell reports

    2017  Volume 18, Issue 8, Page(s) 1840–1847

    Abstract: Endocytosis in neuronal dendrites is known to play a critical role in synaptic transmission and plasticity such as long-term depression (LTD). However, the inability to detect endocytosis directly in living neurons has hampered studies of its dynamics ... ...

    Abstract Endocytosis in neuronal dendrites is known to play a critical role in synaptic transmission and plasticity such as long-term depression (LTD). However, the inability to detect endocytosis directly in living neurons has hampered studies of its dynamics and regulation. Here, we visualized the formation of individual endocytic vesicles containing pHluorin-tagged receptors with high temporal resolution in the dendrites of cultured hippocampal neurons. We show that transferrin receptors (TfRs) are constitutively internalized at optically static clathrin-coated structures. These structures are slightly enriched near synapses that represent preferential sites for the endocytosis of postsynaptic AMPA-type receptors (AMPARs), but not for non-synaptic TfRs. Moreover, the frequency of AMPAR endocytosis events increases after the induction of NMDAR-dependent chemical LTD, but the activity of perisynaptic endocytic zones is not differentially regulated. We conclude that endocytosis is a highly dynamic and stereotyped process that internalizes receptors in precisely localized endocytic zones.
    Language English
    Publishing date 2017-02-21
    Publishing country United States
    Document type Journal Article
    ZDB-ID 2649101-1
    ISSN 2211-1247 ; 2211-1247
    ISSN (online) 2211-1247
    ISSN 2211-1247
    DOI 10.1016/j.celrep.2017.01.081
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  10. Article ; Online: Endocytosis in the axon initial segment maintains neuronal polarity.

    Eichel, Kelsie / Uenaka, Takeshi / Belapurkar, Vivek / Lu, Rui / Cheng, Shouqiang / Pak, Joseph S / Taylor, Caitlin A / Südhof, Thomas C / Malenka, Robert / Wernig, Marius / Özkan, Engin / Perrais, David / Shen, Kang

    Nature

    2022  Volume 609, Issue 7925, Page(s) 128–135

    Abstract: Neurons are highly polarized cells that face the fundamental challenge of compartmentalizing a vast and diverse repertoire of proteins in order to function ... ...

    Abstract Neurons are highly polarized cells that face the fundamental challenge of compartmentalizing a vast and diverse repertoire of proteins in order to function properly
    MeSH term(s) Animals ; Axon Initial Segment/metabolism ; Caenorhabditis elegans ; Cell Membrane/metabolism ; Cell Polarity ; Dendrites/metabolism ; Diffusion ; Endocytosis ; Endosomes/metabolism ; Humans ; Mice ; Protein Transport ; Proteolysis ; Rats ; Receptors, Cell Surface/metabolism
    Chemical Substances Receptors, Cell Surface
    Language English
    Publishing date 2022-08-17
    Publishing country England
    Document type Journal Article
    ZDB-ID 120714-3
    ISSN 1476-4687 ; 0028-0836
    ISSN (online) 1476-4687
    ISSN 0028-0836
    DOI 10.1038/s41586-022-05074-5
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