N-Cadherin Relocalizes from the Periphery to the Center of the Synapse after Transient Synaptic Stimulation in Hippocampal Neurons

N-cadherin is a cell adhesion molecule which is enriched at synapses. Binding of N-cadherin molecules to each other across the synaptic cleft has been postulated to stabilize adhesion between the presynaptic bouton and the postsynaptic terminal. N-cadherin is also required for activity-induced changes at synapses, including hippocampal long term potentiation and activity-induced spine expansion and stabilization. We hypothesized that these activity-dependent changes might involve changes in N-cadherin localization within synapses. To determine whether synaptic activity changes the localization of N-cadherin, we used structured illumination microscopy, a super-resolution approach which overcomes the conventional resolution limits of light microscopy, to visualize the localization of N-cadherin within synapses of hippocampal neurons. We found that synaptic N-cadherin exhibits a spectrum of localization patterns, ranging from puncta at the periphery of the synapse adjacent to the active zone to an even distribution along the synaptic cleft. Furthermore, the N-cadherin localization pattern within synapses changes during KCl depolarization and after transient synaptic stimulation. During KCl depolarization, N-cadherin relocalizes away from the central region of the synaptic cleft to the periphery of the synapse. In contrast, after transient synaptic stimulation with KCl followed by a period of rest in normal media, fewer synapses have N-cadherin present as puncta at the periphery and more synapses have N-cadherin present more centrally and uniformly along the synapse compared to unstimulated cells. This indicates that transient synaptic stimulation modulates N-cadherin localization within the synapse. These results bring new information to the structural organization and activity-induced changes occurring at synapses, and suggest that N-cadherin relocalization may contribute to activity dependent changes at synapses.

[1]  Laurence Pelletier,et al.  Subdiffraction imaging of centrosomes reveals higher-order organizational features of pericentriolar material , 2012, Nature Cell Biology.

[2]  B. Honig,et al.  Thinking outside the cell: how cadherins drive adhesion. , 2012, Trends in cell biology.

[3]  Stephan J Sigrist,et al.  Optical super-resolution microscopy in neurobiology , 2012, Current Opinion in Neurobiology.

[4]  T. Blanpied,et al.  Lateral organization of the postsynaptic density , 2011, Molecular and Cellular Neuroscience.

[5]  Y. Goda,et al.  Differential control of presynaptic efficacy by postsynaptic N-cadherin and β-catenin , 2011, Nature Neuroscience.

[6]  X. Zhuang,et al.  Superresolution Imaging of Chemical Synapses in the Brain , 2010, Neuron.

[7]  Erik B. Bloss,et al.  Persistence of Coordinated Long-Term Potentiation and Dendritic Spine Enlargement at Mature Hippocampal CA1 Synapses Requires N-Cadherin , 2010, The Journal of Neuroscience.

[8]  T. Dresbach,et al.  Essential cooperation of N-cadherin and neuroligin-1 in the transsynaptic control of vesicle accumulation , 2010, Proceedings of the National Academy of Sciences.

[9]  D. Muller,et al.  N-cadherin mediates plasticity-induced long-term spine stabilization , 2010, The Journal of cell biology.

[10]  Fidel Santamaría,et al.  Quantifying the Effects of Elastic Collisions and Non-Covalent Binding on Glutamate Receptor Trafficking in the Post-Synaptic Density , 2010, PLoS Comput. Biol..

[11]  E. S. Ruthazer,et al.  N‐cadherin prodomain cleavage regulates synapse formation in vivo , 2009, Developmental neurobiology.

[12]  Jyothi Arikkath,et al.  Cadherins and catenins at synapses: roles in synaptogenesis and synaptic plasticity , 2008, Trends in Neurosciences.

[13]  M. Gustafsson,et al.  Three-dimensional resolution doubling in wide-field fluorescence microscopy by structured illumination. , 2008, Biophysical journal.

[14]  R. Huganir,et al.  Coordination of Synaptic Adhesion with Dendritic Spine Remodeling by AF-6 and Kalirin-7 , 2008, The Journal of Neuroscience.

[15]  M. Gustafsson,et al.  Subdiffraction Multicolor Imaging of the Nuclear Periphery with 3D Structured Illumination Microscopy , 2008, Science.

[16]  Xiaobing Chen,et al.  Organization of the core structure of the postsynaptic density , 2008, Proceedings of the National Academy of Sciences.

[17]  T. Sakurai,et al.  Activity-Induced Protocadherin Arcadlin Regulates Dendritic Spine Number by Triggering N-Cadherin Endocytosis via TAO2β and p38 MAP Kinases , 2007, Neuron.

[18]  Wei Lu,et al.  Synaptic Anchorage of AMPA Receptors by Cadherins through Neural Plakophilin-Related Arm Protein–AMPA Receptor-Binding Protein Complexes , 2007, The Journal of Neuroscience.

[19]  Paul Greengard,et al.  Three-Dimensional Architecture of Presynaptic Terminal Cytomatrix , 2007, The Journal of Neuroscience.

[20]  E. Schuman,et al.  Activity-Regulated N-Cadherin Endocytosis , 2007, Neuron.

[21]  M. Passafaro,et al.  Extracellular Interactions between GluR2 and N-Cadherin in Spine Regulation , 2007, Neuron.

[22]  S. Kaech,et al.  Culturing hippocampal neurons , 2006, Nature Protocols.

[23]  F. Cordelières,et al.  A guided tour into subcellular colocalization analysis in light microscopy , 2006, Journal of microscopy.

[24]  Robert Moore,et al.  Development/plasticity/repair N-cadherin Transsynaptically Regulates Short-term Plasticity at Glutamatergic Synapses in Embryonic Stem Cell-derived Neurons the Cell Adhesion Molecule N-cadherin Has Been Proposed to Regulate Synapse Formation in Mammalian Central Neurons. This Is Based on Its Synapti , 2022 .

[25]  R. Huganir,et al.  Regulation of AMPA receptor trafficking by N‐cadherin , 2006, Journal of neurochemistry.

[26]  D. Benson,et al.  Structural basis for developmentally regulated changes in cadherin function at synapses , 2006, The Journal of comparative neurology.

[27]  C. Curcio,et al.  Assembly of the N-cadherin complex during synapse formation involves uncoupling of p120-catenin and association with presenilin 1 , 2005, Molecular and Cellular Neuroscience.

[28]  Y. Hiraoka,et al.  Cadherin activity is required for activity-induced spine remodeling , 2004, The Journal of cell biology.

[29]  Kira E. Poskanzer,et al.  Temporally distinct demands for classic cadherins in synapse formation and maturation , 2004, Molecular and Cellular Neuroscience.

[30]  Stephen J. Smith,et al.  In Vivo Trafficking and Targeting of N-Cadherin to Nascent Presynaptic Terminals , 2004, The Journal of Neuroscience.

[31]  W. Birchmeier,et al.  Role of β-Catenin in Synaptic Vesicle Localization and Presynaptic Assembly , 2003, Neuron.

[32]  R. Nicoll,et al.  AMPA Receptor Trafficking at Excitatory Synapses , 2003, Neuron.

[33]  D. Choquet,et al.  Direct imaging of lateral movements of AMPA receptors inside synapses , 2003, The EMBO journal.

[34]  Michael Loran Dustin,et al.  Neural and Immunological Synaptic Relations , 2002, Science.

[35]  M. Takeichi,et al.  Cadherin Regulates Dendritic Spine Morphogenesis , 2002, Neuron.

[36]  E. Schuman,et al.  Depolarization Drives β-Catenin into Neuronal Spines Promoting Changes in Synaptic Structure and Function , 2002, Neuron.

[37]  Yingming Zhao,et al.  The Presynaptic Particle Web Ultrastructure, Composition, Dissolution, and Reconstitution , 2001, Neuron.

[38]  D. Benson,et al.  Stages of Synapse Development Defined by Dependence on F-Actin , 2001, The Journal of Neuroscience.

[39]  O. Bozdagi,et al.  Increasing Numbers of Synaptic Puncta during Late-Phase LTP N-Cadherin Is Synthesized, Recruited to Synaptic Sites, and Required for Potentiation , 2000, Neuron.

[40]  C. Redies.  Cadherins in the central nervous system , 2000, Progress in Neurobiology.

[41]  M. Gustafsson.  Surpassing the lateral resolution limit by a factor of two using structured illumination microscopy , 2000, Journal of microscopy.

[42]  David R. Colman,et al.  Molecular Modification of N-Cadherin in Response to Synaptic Activity , 2000, Neuron.

[43]  Hidekazu Tanaka,et al.  N-Cadherin Redistribution during Synaptogenesis in Hippocampal Neurons , 1998, The Journal of Neuroscience.

[44]  Chou P Hung,et al.  A Role for the Cadherin Family of Cell Adhesion Molecules in Hippocampal Long-Term Potentiation , 1998, Neuron.

[45]  Josef Spacek,et al.  Three‐dimensional organization of cell adhesion junctions at synapses and dendritic spines in area CA1 of the rat hippocampus , 1998, The Journal of comparative neurology.

[46]  D. Colman.  Neurites, Synapses, and Cadherins Reconciled , 1997, Molecular and Cellular Neuroscience.

[47]  R. Weinberg,et al.  Tangential synaptic distribution of NMDA and AMPA receptors in rat neocortex , 1997, Neuroscience Letters.

[48]  M. Takeichi,et al.  The catenin/cadherin adhesion system is localized in synaptic junctions bordering transmitter release zones , 1996, The Journal of cell biology.

[49]  D. Colman,et al.  A Model for Central Synaptic Junctional Complex Formation Based on the Differential Adhesive Specificities of the Cadherins , 1996, Neuron.

[50]  H Korn,et al.  Activity-dependent deformations of presynaptic grids at central synapses , 1985, Journal of neurocytology.

[51]  M. Takeichi.  The cadherin superfamily in neuronal connections and interactions , 2007, Nature Reviews Neuroscience.

[52]  Z. P. I N C U S,et al.  Comparison of quantitative methods for cell-shape analysis , 2007 .

[53]  W. Birchmeier,et al.  Role of beta-catenin in synaptic vesicle localization and presynaptic assembly. , 2003, Neuron.

[54]  E. Schuman,et al.  Depolarization drives beta-Catenin into neuronal spines promoting changes in synaptic structure and function. , 2002, Neuron.