IBENS - ENS

Regulation of molecular interactions between synaptic components

Sarah Reemers — 2018-01-25T13:36:41Z

Head of Project : Christian Specht
Christian Specht CR Inserm

Single molecule localisation microscopy (SMLM) gives access to spatio-temporal distribution patterns that reflect the molecular interactions in which the proteins engage : we can count the number and density of molecules in specific compartments, visualise their localisation with high spatial precision, and track their movements at high temporal resolution in living cells. The comparison with biochemical approaches shows that the mobility of cellular components is inversely related to the binding constants of purified proteins, demonstrating that SMLM data can yield information about molecular interactions in living cells.

Changes in the number or the binding properties of synaptic proteins necessarily alter the organisation of the synaptic structure, which in turn affects the efficacy of synaptic transmission. Using SMLM strategies our project explores the interactions between membrane receptors, scaffold and adhesion proteins as well as signalling and cytoskeletal elements at synapses. We investigate how the binding properties of different pre- and postsynaptic components are regulated through network activity and signalling processes in cultured neurons and organotypic slices.

Participants : C. Specht, X. Yang, A. Jan, F. Niwa, and N. Frézel

Grünewald N, Jan A, Salvatico C, Kress V, Renner R, Triller A, Specht CG, Schwarz G (2018). Sequences flanking the gephyrin-binding site of GlyR-beta tune receptor stabilization at synapses. eNeuro.

Patrizio A, Renner M, Pizzarelli R, Triller A, Specht CG (2017). Alpha subunit-dependent glycine receptor clustering and regulation of synaptic receptor numbers. Sci Rep 7:10899.

Salvatico C, Specht CG and Triller A (2015). Synaptic receptor dynamics : from theoretical concepts to deep quantification and chemistry in cellulo. Neuropharmacol 88:2-9.

Specht CG, Izeddin I, Rodriguez PC, El Beheiry M, Rostaing P, Darzacq X, Dahan M and Triller A (2013). Quantitative nanoscopy of inhibitory synapses : counting gephyrin molecules and receptor binding sites. Neuron 79:308-321.

Inhibitory synapses regulation by glia and inflammation

Andrea Dumoulin (équipe Triller) — 2015-02-11T14:00:47Z

Head of project : Alain Bessis
Alain Bessis DR CNRS

Microglia are non-neuronal cells of the nervous tissue. They display strong phagocytic activity, and the microglial functions that have been best described are related to their ability eliminate dead cells, cellular debris etc. However, it is more and more accepted that microglia are more than “housekeeping cells” but rather are full-time partners of neuronal function.

Thus, microglia are extremely ramified and dynamic cells that permanently extend and retract their processes to scan the brain and rapidly react to the modification of the environment. We and others have shown that microglia are able to induce the physiological death of developing neurons and that they participate to the growth of axons during development.

We showed that microglia are genuine partners of excitatory neurotransmission and we are now investigating how microglia can tune the synaptic function by controlling the behaviour of neurotransmitter receptors. To this aim, we are using up-to-date imaging approaches to monitor receptors dynamic and microglia-synapses interactions.

Participants : A. Bessis, I. Vaccari, Y. Cantaut-Belarif, S. Colasse

Cantaut-Belarif Y, M. Antri, R. Pizzarelli, S. Colasse, I. Vaccari, S. Soares, M. Renner, R. Dallel, A. Triller, and A. Bessis (2017) Microglia control the glycinergic but not the GABAergic synapses via prostaglandin E2 in the spinal cord. The Journal of Cell Biology. jcb.201607048. doi:10.1083/jcb.201607048

Béchade C, S. Colasse, M. Diana, M. Rouault, and A. Bessis (2014) NOS2 expression is restricted to neurons in the healthy brain but is triggered in microglia upon inflammation. Glia. 62:956–63. doi:10.1002/glia.22652

Squarzoni P, G. Oller, G. Hoeffel, L. Pont-Lezica, P. Rostaing, D. Low, A. Bessis, F. Ginhoux, and S. Garel (2014) Microglia modulate wiring of the embryonic forebrain. CellReports. 8:1271-9. doi:10.1016/j.celrep. 2014.07.042

Pont-Lezica L, W. Beumer, S. Colasse, H. Drexhage, M. Versnel, and A. Bessis (2014) Microglia shape corpus callosum axon tract fasciculation : functional impact of prenatal inflammation. Eur J Neurosci. 39:1551–7. doi:10.1111/ejn.12508

Pascual O, S.B. Achour, P. Rostaing, A. Triller, and A. Bessis (2012) Microglia activation triggers astrocyte-mediated modulation of excitatory neurotransmission. Proc Natl Acad Sci USA. 109:E197–205. doi:10.1073/ pnas.1111098109

Fundamental mechanisms and human-specific regulations of synaptic development and plasticity

Andrea Dumoulin (équipe Triller) — 2015-02-11T13:56:52Z

Head of Project : Cécile Charrier

Synapses are multimolecular nanomachines that ensure the proper connectivity of neuronal circuits, integrate diverse inputs into biochemical reactions and allow adaptive responses to the environment. Small changes in the molecular organization of synapses can lead to profound modifications of behavioral and cognitive abilities, as frequently observed in neurodevelopmental and psychiatric disorders. Small changes also appeared during human evolution. Although poorly characterized, they underlie the distinctive developmental and physiological properties of human synapses and are at the core of what makes us humans.

We are investigating the role of molecular pathways linked to human evolution in the development and plasticity of synaptic connections in the neocortex. Our goal is to elucidate fundamental principles that are common to all mammals, and uncover regulations that are specific to humans. We employ a multidisciplinary approach in mouse and human models based on sparse in vivo manipulations in intact cortical circuits, proteomics, electrophysiology, as well as confocal, super-resolutive, and correlative microscopy.

Our recent work has focused on SRGAP2, one of the few genes specifically duplicated in the human lineage. We have demonstrated that the human-specific copy of SRGAP2 inhibits the ancestral protein, which leads to the emergence of human features of synapses such as their protracted developmental period and their increased density along dendrites. By investigating the underlying molecular basis, we have unraveled an intrinsic mechanism coordinating the development of excitatory and inhibitory synapses.

Participants : C. Charrier, M. Fossati, N. Assendorp, O. Gemin, S. Colasse and P. Rostaing.

Funding : ANR, Fyssen Foundation

Maffei A, Charrier C, Caiati MD, Barberis A, Mahadevan V, Woodin MA, Tyagarajan SK (2017) Emerging Mechanisms Underlying Dynamics of GABAergic Synapses. J Neurosci. 37:10792-10799

Fossati M, Pizzarelli R, Schmidt ER, Kupferman JV, Stroebel D, Polleux F, Charrier C (2016) SRGAP2 and Its Human-Specific Paralog Co-Regulate the Development of Excitatory and Inhibitory Synapses. Neuron 91:356-69

Charrier C, Polleux F (2012) [How human-specific SRGAP2 gene duplications control human brain development]. Med Sci (Paris) 28:911-4

Charrier C, Joshi K, Coutinho-Budd J, Kim JE, Lambert N, de Marchena J, Jin WL, Vanderhaeghen P, Ghosh A, Sassa T, Polleux F (2012) Inhibition of SRGAP2 function by its human-specific paralogs induces neoteny during spine maturation. Cell 149:923-35

Synaptic microstructure, dynamics and molecular interactions

Andrea Dumoulin (équipe Triller) — 2015-02-11T11:40:21Z

Three directions are considered here, and through their combination we aim at a deeper understanding of the molecular dynamics at synapses.

1- The transmission of signals between neurons is a function of the molecular organisation of synapses. In other words, the function of a synapse is determined by the number, activity and precise location of its components. In order to explore the ultrastructure of excitatory and inhibitory synapses we use advanced electron microscopy techniques such as high pressure freezing (HPF) and correlative light and electron microscopy (CLEM). In addition, we have implemented photo-activated localisation microscopy (PALM) to elucidate the absolute numbers and distribution of synaptic proteins at super-resolution scale.

Participants : A. Triller, P. Rostaing, F. Niwa, A. Ludwig, P. Serna Martinez, O. Gemin

2- Functional changes at synapses are necessarily the result of dynamic rearrangements of the synaptic structure. Over the last decade our laboratory has investigated the diffusion properties of inhibitory neurotransmitter receptors using single particle tracking (SPT) of quantum dot (QD) tagged receptor subunits at high spatial and temporal resolution. These studies have revealed the dynamic exchange of receptors between the synaptic and the extra-synaptic membrane as well as their immobilisation at synapses. More recently, we have begun to investigate the molecular interactions between receptors and scaffold proteins at synapses using high-density PALM-based single molecule imaging.

Participants : C. Specht, A. Triller, X. Yang, A. Jan, S. Colasse, P. Rostaing

3- Dynamic rearrangements of the synaptic structure are the basis of synaptic plasticity. Using pharmacological and physiological approaches as well as recombinant probes, we explore the upstream signalling processes that regulate the structure and function of synapses in cultured neurons. This includes changes in synaptic activity, activation of signalling molecules and post-translational modifications, the role of the extracellular matrix (ECM) and the presence of diffusion barriers for the dynamic exchange of synaptic components.

Participants : A. Dumoulin, A. Triller, V. Lepetz, P. Rostaing, S. Colasse

Patrizio A, Renner M, Pizzarelli R, Triller A, Specht CG (2017) Alpha subunit-dependent GlyR clustering and regulation of synaptic occupancy. Scientific Reports In press

Renner M, Wang L, Levi S, Hennekinne L, Triller A (2017) A simple and powerful analysis of lateral subdiffusion using single particle tracking. Biophys J in press

Shrivastava AN, Aperia A, Melki R, Triller A (2017) Physico-Pathologic Mechanisms involved in Neurodegeneration : Misfolded Proteins-Plasma Membrane Interactions. Neuron 95(1):33-50 ; doi : 10.1016

Bannai H, Niwa F, Sherwood MW, Shrivastava AN, Arizono M, Miyamoto A, Sugiura K, Lévi S, Triller A, Mikoshiba K (2015) Bidirectional Control of Synaptic GABAAR Clustering by Glutamate and Calcium. Cell Report 13:2768-80

Shrivastava AN, Redeker V, Fritz N, Pieri L, Almeida LG, Spolidoro M, Liebmann T, Bousset L, Renner M, Léna C, Aperia A, Melki R, Triller A (2015) α-synuclein assemblies sequester neuronal 3-Na+/K+-ATPase and impair Na+ gradient. EMBO J 34:2408-23

Specht CG, Izeddin I, Rodriguez PC, El Beheiry M, Rostaing P, Darzacq X, Dahan M, Triller A (2013) Quantitative nanoscopy of inhibitory synapses : counting gephyrin molecules and receptor binding sites. Neuron 79 : 308-321

Publications

Andrea Dumoulin (équipe Triller), Sarah Reemers — 2015-02-10T13:55:08Z

2017

Mukherjee J, Cardarelli RA, Cantaut-Belarif Y, Deeb TZ, Srivastava DP, Tyagarajan SK, Pangalos MN, Triller A, Maguire J, Brandon NJ, Moss SJ. (2017) Estradiol modulates the efficacy of synaptic inhibition by decreasing the dwell time of GABAA receptors at inhibitory synapses. Proc Natl Acad Sci U S A 114:11763-11768

Patrizio A, Renner M, Pizzarelli R, Triller A, Specht CG (2017) Alpha subunit-dependent GlyR clustering and regulation of synaptic occupancy. Scientific Reports In press

Renner M, Wang L, Levi S, Hennekinne L, Triller A (2017) A simple and powerful analysis of lateral subdiffusion using single particle tracking. Biophys J in press

Cantaut-Belarif Y, Antri M, Pizzarelli R, ColasseS, Vaccari I, Soares S, Renner M, Dallel R, Triller A*, Bessis B* (2017) Microglia control the glycinergic but not the GABAergic synapses via prostaglandin E2 in the spinal cord J Cell Biol 216:2979-2989

Ranft J, Almeida LG , Rodriguez PC , Triller A*, HakimV* (2017) An aggregation-removal model for the formation and size determination of post-synaptic scaffold domains. PloS Comput Biology Biol. 2017 Apr 24 ;13(4):e1005516

Shrivastava AN, Aperia A, Melki R, Triller A (2017) Physico-Pathologic Mechanisms involved in Neurodegeneration : Misfolded Proteins-Plasma Membrane Interactions. Neuron 95(1):33-50 ; doi : 10.1016

Maffei A, Charrier C, Caiati MD, Barberis A, Mahadevan V, Woodin MA, Tyagarajan SK (2017) Emerging Mechanisms Underlying Dynamics of GABAergic Synapses. J Neurosci. 37:10792-10799

2016

Hannan S, Gerrow K, Triller A, Smart TG (2016) Phospho-dependent Accumulation of GABABRs at Presynaptic Terminals after NMDAR Activation. Cell Rep. 16:1962-73

Shrivastava AN, Redeker V, Fritz N, Pieri L, Almeida LG, Spolidoro M, Liebmann T, Bousset L, Renner M, Léna C, Aperia A, Melki R, Triller A (2016) Data in support of the identification of neuronal and astrocyte proteins interacting with extracellularly applied oligomeric and fibrillar synuclein assemblies by mass spectrometry. Data in Brief 7 : 221-8

El Beheiry M, T¨urkcan S, Richly M, Triller A, Alexandrou A, Dahan M & Masson J.-B. (2016) A Primer on the Bayesian Approach to High-density Single-Molecule Trajectories Analysis. Biophysical J 110:1209-15

Wang I, Dumoulin A, Renner M, Triller A*, Specht CG (2016) The Role of Synaptopodin in Membrane Protein Diffusion in the Dendritic Spine Neck PLoS ONE 11(2) : 1-18 e0148310. doi:10.1371/

2015

Plamont MA, Billon-Denis E, Maurin S, Gauron C, Pimenta FM, Specht CG, Shi J, Quérard J, Pan B, Rossignol J, Morellet N, Volovitch M, Lescop E, Chen Y, Triller A, Vriz S, Le Saux T, Jullien L, Gautier A (2015) A small fluorescence-activating and absorption-shifting tag for tunable protein imaging in vivo. Proc Natl Acad Sci USA 113:497-502

Haselwandter CA, Kardar M, Triller A, da Silveira RA (2015) Self-assembly and plasticity of synaptic domains through a reaction-diffusion mechanism. Phys Rev E Stat Nonlin Soft Matter Phys. 92(3-1):032705. Epub 2015 Sep 3

Hausrat T, Muhia M, Gerrow K, Thomas P, Hirdes W, Tsukita S, Heisler F, Herich L, Dubroqua S, Breiden P, Feldon J, Schwarz J, Yee B, Smart T, Triller A, Kneussel M (2015) Radixin regulates synaptic GABAA receptor density and is essential for reversal learning and short-term memory. Nature Com. doi:10.1038/ncomms7872

Nadjar Y, Triller A, Bessereau JL, Dumoulin A (2015) The Susd2 protein regulates neurite growth and excitatory synaptic density in hippocampal cultures. Mol Cell Neurosci. 65:82-91

Salvatico C, Specht CG, Triller A (2015) Synaptic receptor dynamics : from theoretical concepts to deep quantification and chemistry in cellulo. Neuropharmacology. 88:2-9

2014

Gouzer G, Specht CG, Allain L, Shinoe T, Triller A (2014) Benzodiazepine-dependent stabilization of GABA(A) receptors at synapses. Mol Cell Neurosci. 63:101-13

Kneussel M, Triller A, Choquet D (2014) SnapShot : receptor dynamics at plastic synapses.
Cell. 157(7):1738-1738.e1

Gerrow K, Triller A. (2014) GABAA receptor subunit composition and competition at synapses are tuned by GABAB receptor activity. Mol Cell Neurosci. 60:97-107

Masson JB, Dionne P, Salvatico C, Renner M, Specht CG, Triller A, Dahan M (2014) Mapping the energy and diffusion landscapes of membrane proteins at the cell surface using high-density single-molecule imaging and Bayesian inference : application to the multiscale dynamics of glycine receptors in the neuronal membrane. Biophys J 106 : 74-83

Horellou S, Pascual O, Triller A, Marty S (2014) Adaptive and non-adaptive changes in activity-deprived presynaptic terminals. Eur J Neurosci 39 : 61-71

2013

Siksou L, Silm K, Biesemann C, Nehring RB, Wojcik SM, Triller A, El Mestikawy S, Marty S, Herzog E (2013) A role for vesicular glutamate transporter 1 in synaptic vesicle clustering and mobility. Eur J Neurosci 37 : 1631-1642

Shrivastava AN, Rodriguez PC, Triller A, Renner M (2013) Dynamic micro-organization of P2X7 receptors revealed by PALM based single particle tracking. Front Cell Neurosci 7(232) : doi : 10.3389

Shrivastava AN, Kowalewski JM, Renner M, Bousset L, Koulakoff A, Melki R, Giaume C, Triller A, (2013) beta-amyloid and ATP-induced diffusional trapping of astrocyte and neuronal metabotropic glutamate type-5 receptors. Glia 61 : 1673-1686

Hennekinne L, Colasse S, Triller A, Renner M (2013) Differential control of thrombospondin over synaptic glycine and AMPA receptors in spinal cord neurons. J Neurosci 33 : 11432-11439

Choquet D, Triller A (2013) The dynamic synapse. Neuron 80 : 691-703

2012

Triller A, Sheng M (2012) Synaptic structure and function. Curr Opin Neurobiol 22 : 363-365

Renner M, Schweizer C, Bannai H, Triller A, Levi S (2012) Diffusion barriers constrain receptors at synapses. PLoS One 7 : e43032

Pascual O, Ben Achour S, Rostaing P, Triller A, Bessis A (2012) Microglia activation triggers astrocyte-mediated modulation of excitatory neurotransmission. Proc Natl Acad Sci U S A 109 : E197-E205

Niwa F, Bannai H, Arizono M, Fukatsu K, Triller A, Mikoshiba K (2012) Gephyrin-Independent GABA(A)R Mobility and Clustering during Plasticity. PLoS One 7 : doi/10.1371

Izeddin I, Boulanger J, Racine V, Specht CG, Kechkar A, Nair D, Triller A, Choquet D, Dahan M, Sibarita JB (2012) Wavelet analysis for single molecule localization microscopy. Opt Express 20 : 2081-2095

2011

Triller A (2011f) Molecular Imagery : towards in cellulo chemistry. MS-Med Sci 27 : 451-452

Specht CG, Grunewald N, Pascual O, Rostgaard N, Schwarz G, Triller A (2011) Regulation of glycine receptor diffusion properties and gephyrin interactions by protein kinase C. Embo J 30 : 3842-3853

Sillibourne JE, Specht CG, Izeddin I, Hurbain I, Tran P, Triller A, Darzacq X, Dahan M, Bornens M (2011) Assessing the localization of centrosomal proteins by PALM/STORM nanoscopy. Cytoskeleton 68) : 619-627

Siksou L, Triller A, Marty S (2011) Ultrastructural organization of presynaptic terminals. Curr Opin Neurobiol 21 : 261-268

Shrivastava AN, Triller A, Sieghart W, Sarto-Jackson I (2011) Regulation of GABA(A) Receptor Dynamics by Interaction with Purinergic P2X(2) Receptors. J Biol Chem 286 : 14455-14468

Shrivastava AN, Triller A, Sieghart W (2011) GABA(A) Receptors : Post-Synaptic Co-Localization and Cross-Talk with Other Receptors. Front Cell Neurosci 5 : doi : 10.3389/fncel.2011.00007

Ribrault C, Sekimoto K, Triller A (2011) From the stochasticity of molecular processes to the variability of synaptic transmission. Nature Rev Neurosci 12 : 375-387

Ribrault C, Reingruber J, Petkovic M, Galli T, Ziv NE, Holcman D, Triller A (2011) Syntaxin1A lateral diffusion reveals transient and local SNARE interactions. J Neurosci 31 : 17590-17602

Renner M, Domanov Y, Sandrin F, Izeddin I, Bassereau P, Triller A (2011) Lateral diffusion on tubular membranes : quantification of measurements bias. PLoS One 6 : e25731

Mukherjee J, Kretschmannova K, Gouzer G, Maric HM, Ramsden S, Tretter V, Harvey K, Davies PA, Triller A, Schindelin H, Moss SJ (2011) The Residence Time of GABA(A)Rs at Inhibitory Synapses Is Determined by Direct Binding of the Receptor alpha 1 Subunit to Gephyrin. J Neurosci 31 : 14677-14687

Machado P, Rostaing P, Guigonis JM, Renner M, Dumoulin A, Samson M, Vannier C, Triller A (2011) Heat Shock Cognate Protein 70 Regulates Gephyrin Clustering. J Neurosci 31 : 3-14

Levi S, Dahan M, Triller A (2011) Labeling neuronal membrane receptors with quantum dots. Cold Spring Harb Protoc 2011 (1) : prot5580

Izeddin I, Specht CG, Lelek M, Darzacq X, Triller A, Zimmer C, Dahan M (2011) Super-Resolution Dynamic Imaging of Dendritic Spines Using a Low-Affinity Photoconvertible Actin Probe. PLoS One 6 : e15611

Haselwandter CA, Calamai M, Kardar M, Triller A, da Silveira RA (2011) Formation and Stability of Synaptic Receptor Domains. Phys Rev Lett 106 : 238104

Domanov YA, Aimon S, Toombes GES, Renner M, Quemeneur F, Triller A, Turner MS, Bassereau P (2011) Mobility in geometrically confined membranes. Proc Natl Acad Sci U S A 108 : 12605-12610

Bechade C, Pascual O, Triller A, Bessis A (2011) Nitric oxide regulates astrocyte maturation in the hippocampus : Involvement of NOS2. Molec Cell Neurosci 46 : 762-769

Thèmes de recherche

Andrea Dumoulin (équipe Triller), Sarah Reemers — 2015-02-10T13:54:07Z

Synaptic microstructure, dynamics and molecular interactions

Humanization of synapse development and plasticity

Regulation of molecular interactions between synaptic components

Inhibitory synapses regulation by glia and inflammation

Membres de l'équipe

Andrea Dumoulin (équipe Triller), Sarah Reemers — 2014-07-02T10:05:03Z

Antoine Triller DR Inserm
Alain Bessis DR CNRS	Andréa Dumoulin MC ENS	Christian Specht CR Inserm

ITAs

Nadège D'Araujo TR Inserm	Sabrina Colasse AI Inserm
Philippe Rostaing IR1 Inserm	Hervé Gendrot TR Inserm

Post-doctorant(e)s

Matteo Fossati	Anastasia Ludwig	Fumihiro Niwa
Maria Pinto	Pablo Serna Martinez

Doctorant(e)s

Nora Assendorp	Olivier Gemin	Audric Jan
Valentin Lepetz	Xiaojuan Yang

Anciens membres

Cécile Charrier (CR Inserm) now IBENS Paris
Serge Marty (CR CNRS) now PI, ICM Paris
Catherine Béchade (CR Inserm) now Institut du Fer à Moulin, Paris
Sabine Lévi (CR CNRS) now Institut du Fer à Moulin, Paris
Lorena Pont-Lezica (Postdoc) now Project manager, Essilor International
Kimberly Gerrow (Postdoc) now Postdoc, University of Victoria, Canada
Shirley Wakselman (Postdoc) now Regional Start-Up Lead II, PRA Health Sciences, Paris
Damien Alcor (Postdoc) now Responsable plateforme imagerie, IBDM
Anne Roumier (Postdoc) now PI Institut du Fer à Moulin, Paris and Assistant Professor, Sorbonne Université
Hiroko Bannai (Postdoc) now PI, RIKEN BSI, Saitama, Japan
Marianne Renner (Postdoc) now Professor, Paris UPMC
Pamela RODRIQUEZ (Postdoc) now at Alexion RD France (Imagine)
Leandro ALMEIDA (Postdoc) now at Keen Eye Tech Co (Start-Up)
Cyrille Sur (PhD) now Executive Director, Neuroscience Imaging Lead, Merck & Co
Süzel Horellou (PhD) now Chargée de mission Transferts de Technologies, Région IDF
Géraldine Gouzer (PhD) now Project Engineer, Office of Scientific and Medical Affairs, ICM Paris
Léa Siksou (PhD) now Postdoc, Friedrich Miescher Institute, Basel, Switzerland
Patricia Machado (PhD) now Laboratoires Servier, France
Yann Nadjar (PhD) now Chef de Clinique, Hôpital Pitié-Salpêtrière, Paris
Delphine Bernard (PhD) now Research Project Leader, MedDay Pharmaceuticals, Paris
Lydia Danglot (PhD) now PI, Center for Psychiatry and Neurosciences, Paris
Cyril Hanus (PhD) now Group leader, Center for Psychiatry and Neurosciences, Paris
Charlotte Salvatico (PhD), now movie producer, Science Mediation
Laetitia Hennekine (PhD), now sister in the Carmelites, Spain