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Membrane Traffic at Synapses

Team Leader : David Perrais

David Perrais biosketch

The team (July 2020)

From left to right (without masks): Julie, Lou, Etienne,

David, Silvia, Vincent, May, Zehra, Mathias

 

General objective

Vesicular trafficking is one of the most salient features of synaptic physiology. In the tiny (less than 1 µm wide) chemical synapses, presynaptic vesicles concentrate and release neurotransmitter molecules which bind to post-synaptic receptors. The exocytosis and recycling of synaptic vesicles is a very prominent and essential feature of neuronal physiology that is highly controlled in time and space. Moreover, post-synaptic membrane trafficking, although not as prominent quantitatively, is pivotal for the maintenance of signal transduction complexes supporting synaptic transmission and plasticity. Most of our knowledge about synapse physiology comes from studying glutamatergic synapses which represent the majority of synapses in the brain. Nevertheless, other types of synapses, such as neuromodulatory dopaminergic synapses, could have a very different molecular composition and operate in a different way. However, because they represent a small minority of synapses formed from a very small number of neurons, their analysis has been difficult through classical cellular and molecular methods.

Our goal in the team is to use the most advanced fluorescence imaging techniques together with refined purification of synaptic elements (synaptosomes) to address the mechanisms regulating synapse function through membrane trafficking events in normal brain physiology or in the course of disease. To achieve this goal, we use, on top of the standard techniques of the modern neuroscience lab (molecular biology, biochemistry, imaging, electrophysiology), two unique expertise developed by the two PIs: first, with David Perrais, we develop methods to detect individual exocytosis and endocytosis events with pH sensitive fluorophores and perform quantitative imaging. Second, with Etienne Herzog, we purify synaptosomes from adult animals with fluorescence activated synaptosome sorting (FASS), which enables powerful proteomics, transcriptomics and functional approaches.

Altogether, we aim at identifying new pathways in specific synapses and test their relevance for synaptic nanostructure and function in the normal and diseased brain.

Research Projects

Local Regulation of receptor trafficking in dendrites

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Molecular and cellular mechanisms of endocytosis

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Synaptomic dissection of Dopaminergic projections

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Synaptic alterations and therapeutic approaches in neurodegenerative diseases

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Expertise

  • Imaging endocytosis with the ppH protocol
  • Ex-vivo cellular and molecular analysis of synapses
  • Semi-quantitative imaging of synaptic vesicle super-pool
  • News

    Post-synaptic exocytosis and synaptic plasticity - Cell Reports Sept 2021

    Synapses, the basic building blocks of neural networks, are both very stable and capable of rapid and long-lasting modifications, a phenomenon known as synaptic plasticity. The modification of a synapse often involves the addition of synaptic receptors (long-term potentiation or LTP) or the removal of part of the synaptic receptors (long-term depression or LTD). This rapid plasticity is possible because synaptic receptors are not immobile in the synapse but travel to intracellular compartments called recycling endosomes (RE). The regulation of RE trafficking has thus become an important topic of study for understanding the mechanisms of synaptic plasticity.

    Picture of all contributors of this study. David Perrais and May Bakr stand in front of "The paint pipette", taken by May, which was awarded the best IINS picture prize in 2020.


    In this study, we searched for the molecules involved in these phenomena, in particular the proteins responsible for exocytosis called SNAREs. The VAMP2 protein, target of the tetanus toxin (released by the bacterium responsible for tetanus and one of the most deadly in humans), was known to block LTP. However, to our surprise, it only marginally affected RE exocytosis. We therefore searched for other SNARE proteins and found VAMP4 to be responsible for the majority of RE exocytosis, whereas VAMP2 is involved in only a small fraction of exocytosis, but plays a major role in the exocytosis of REs containing AMPA-type postsynaptic receptors (see Figure). Furthermore, VAMP4 deletion also alters the trafficking of AMPA receptors that are in greater quantity at the surface of neurons, increasing synaptic transmission and limiting its plasticity by occlusion.
    This work shows the great diversity of membrane trafficking mechanisms in the dendrites of neurons that allows receptors to be delivered when and where they are needed to regulate individual synapses. It was the result of a long-term work, over more than eight years, of many students, engineers and researchers of the IINS.

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    Selected Publications

  • May Bakr, Damien Jullié, Julia Krapivkina, Vincent Paget-Blanc, Lou Bouit, Jennifer Petersen, Natacha Retailleau, Christelle Breillat, Etienne Herzog, Daniel Choquet, David Perrais
  • The vSNAREs VAMP2 and VAMP4 control recycling and intracellular sorting of post-synaptic receptors in neuronal dendrites Cell Reports (2021)
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  • Silvia Sposini, Morgane Rosendale, Léa Claverie, TN Ngoc Van, Damien Jullié & David Perrais
  • Imaging endocytic vesicle formation at high spatial and temporal resolutions with the pulsed-pH protocol Nature Protocols (2020)
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  • Morgane Rosendale, TN Ngoc Van, Dolors Grillo-Bosch, Silvia Sposini, Léa Claverie, Isabel Gautereau, Stéphane Claverol, Daniel Choquet, Matthieu Sainlos & David Perrais
  • Functional recruitment of dynamin requires multimeric interactions for efficient endocytosis Nature Communications (2019)
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  • Hafner A-S, Donlin-Asp PG, Leitch B, Herzog E & Schuman EM.
  • Local protein synthesis is a ubiquitous feature of neuronal pre- and postsynaptic compartments. Science (2019)
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  • Blumenstock S, angelo MF, Peters F, Dorostkar MM, Ruf VC, Luckner M, Crux S, Slapakova L, Arzberger T, Claverol S, Herzog E* & Herms J*
  • Early defects in translation elongation factor 1α levels at excitatory synapses in α-synucleinopathy Acta Neuropathologica (2019)
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  • Zhang XM, Francois U, Silm K, angelo MF, Fernández Busch MV, Maged M, Martin C, Cordelières FP, Deshors M, Pons S, Maskos U, Bemelmans A-P, Wojcik SM, Mestikawy El S, Humeau Y & Herzog E
  • A proline-rich motif on VGLUT1 reduces synaptic vesicle super-pool and spontaneous release frequency eLife (2019)
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  • Magalie Martineau, Agila Somasundaram, Jonathan B Grimm, Todd D Gruber, Daniel Choquet, Justin W Taraska, Luke D Lavis & David Perrais
  • Semisynthetic fluorescent pH sensors for imaging exocytosis and endocytosis Nature Communications (2017)
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  • Morgane Rosendale, Damien Jullié, Daniel Choquet & David Perrais
  • Spatial and temporal regulation of receptor endocytosis in neuronal dendrites revealed by imaging of single vesicle formation Cell Reports (2017)
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    Members

    « Researcher »

    HERZOG Etienne Researcher etienne.herzog@u-bordeaux.fr +33533514861
    PERRAIS David Researcher david.perrais@u-bordeaux.fr +33533514861

    « Technical Staff »

    ANGIBAUD Julie Technical staff julie.angibaud@u-bordeaux.fr +33533514748

    « Postdoc »

    BELAPURKAR Vivek Postdoc vivek.belapurkar@u-bordeaux.fr +33533514700
    JELINKOVA Sarka Postdoc sarka.jelinkova@u-bordeaux.fr +33533514861
    PAGET-BLANC Vincent Postdoc vincent.paget-blanc@u-bordeaux.fr +33533514861
    SPOSINI Silvia Postdoc silvia.sposini@u-bordeaux.fr +33533514861

    « PhD student »

    LAPIOS Paul PhD student paul.lapios@u-bordeaux.fr +33533514779

    « Alumni & Guests »

    Anne-Sophie Hafner (Guest, 2017-2018)

    Magalie Martineau (post-doc, 2015-2019 - Now  Research and Development specialist at EIF Innovation, Paris, France)

    Léa Claverie (PhD student, 2015-2019 - Now Project Manager at Euroquality, Bordeaux, France)

    Florencia Angelo (Lab Manager, 2015-2020)

    Thi Nhu Ngoc Van (post-doc, 2014-2016 - Now Project Manager at Sys2dia, Montpellier, France)

    Julia Krapivkina (PhD student, 2012-2016 - Now Project Manager at Assystem, Paris, France)

    Xiao Min Zhang (PhD Student, 2012-2016 - Now Assistant Prof at Sun Yat Sen University - China)

    Morgane Rosendale (PhD student and post-doc, 2011-2016 - Now post-doctoral fellow at Institute for Molecular Sciences, Bordeaux)

    Damien Jullié (PhD student, 2008-2012 - Now Research Specialist at UCSF Weill Institute for Neurosciences, San Francisco, USA)

    Fundings

    ANR LocalEndoProbes
    ANR-PRC 2019 DopamineHub
    CoEN Pathfinder IV 2019 - LocalDementia