Postsynaptic protein mobility in dendritic spines: Long-term regulation by synaptic NMDA receptor activation

Reorganization of molecular components represents a cellular mechanism for synaptic plasticity. Dendritic spines, major sites for glutamatergic synapses, compartmentalize dynamic changes in molecular composition. Here, we use fluorescence recovery after photobleaching (FRAP) in cultured hippocampal neurons to show that spine proteins undergo continual exchange with extra-spine pools. Each spine component has a distinctive mobility: calcium/calmodulin activated protein kinase CaMKIIalpha > GluR1 AMPA glutamate receptor > PSD-95 scaffolding protein > NR1 NMDA glutamate receptor. Stimulation of synaptic NMDA receptors by a protocol that induces chemical LTP resulted in a long-lasting reduction in the mobility of spine CaMKIIalpha and an increased mobile fraction but slower kinetics for spine GluR1. Stimulation also increased the resistance of postsynaptic CaMKIIalpha to detergent extraction. These results suggest long-lasting changes in affinity of protein-protein interactions and/or ongoing alterations in exo/endocytosis. Such lasting changes in protein mobility may contribute to maintaining alterations in synaptic efficacy.

[1]  H. Schulman,et al.  Regulation of signal transduction by protein targeting: the case for CaMKII. , 2001, Biochemical and biophysical research communications.

[2]  Dominique Muller,et al.  Increased Phosphorylation of Ca/Calmodulin-dependent Protein Kinase II and Its Endogenous Substrates in the Induction of Long Term Potentiation (*) , 1995, The Journal of Biological Chemistry.

[3]  G. Banker,et al.  Culturing nerve cells , 1998 .

[4]  K. Sobue,et al.  Rapid Redistribution of the Postsynaptic Density Protein PSD-Zip45 (Homer 1c) and Its Differential Regulation by NMDA Receptors and Calcium Channels , 2001, The Journal of Neuroscience.

[5]  M. Fischer,et al.  Rapid Actin-Based Plasticity in Dendritic Spines , 1998, Neuron.

[6]  A. Craig,et al.  Postsynaptic Scaffolds of Excitatory and Inhibitory Synapses in Hippocampal Neurons: Maintenance of Core Components Independent of Actin Filaments and Microtubules , 2000, The Journal of Neuroscience.

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

[8]  P. H. Seeburg,et al.  Spatial memory dissociations in mice lacking GluR1 , 2002, Nature Neuroscience.

[9]  John Lisman,et al.  Persistent Accumulation of Calcium/Calmodulin-Dependent Protein Kinase II in Dendritic Spines after Induction of NMDA Receptor-Dependent Chemical Long-Term Potentiation , 2004, The Journal of Neuroscience.

[10]  G. Ellis‐Davies,et al.  Structural basis of long-term potentiation in single dendritic spines , 2004, Nature.

[11]  Xiaobing Chen,et al.  Distribution of Postsynaptic Density (PSD)-95 and Ca2+/Calmodulin-Dependent Protein Kinase II at the PSD , 2003, The Journal of Neuroscience.

[12]  Guo-Qiang Bi,et al.  Spatiotemporal specificity of synaptic plasticity: cellular rules and mechanisms , 2002, Biological Cybernetics.

[13]  T Suzuki,et al.  Rapid Translocation of Cytosolic Ca2+/Calmodulin‐Dependent Protein Kinase II into Postsynaptic Density After Decapitation , 1994, Journal of neurochemistry.

[14]  R. Malenka,et al.  AMPA receptor trafficking and synaptic plasticity. , 2002, Annual review of neuroscience.

[15]  G. Westbrook,et al.  Mobile NMDA Receptors at Hippocampal Synapses , 2002, Neuron.

[16]  J. Lippincott-Schwartz,et al.  Studying protein dynamics in living cells , 2001, Nature Reviews Molecular Cell Biology.

[17]  N. Demaurex.  pH Homeostasis of cellular organelles. , 2002, News in physiological sciences : an international journal of physiology produced jointly by the International Union of Physiological Sciences and the American Physiological Society.

[18]  A. Craig,et al.  Rapid Synaptic Remodeling by Protein Kinase C: Reciprocal Translocation of NMDA Receptors and Calcium/Calmodulin-Dependent Kinase II , 2002, The Journal of Neuroscience.

[19]  A. Triller,et al.  The role of receptor diffusion in the organization of the postsynaptic membrane , 2003, Nature Reviews Neuroscience.

[20]  C. Winters,et al.  Glutamate-induced transient modification of the postsynaptic density , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[21]  Venkatesh N. Murthy,et al.  Rapid turnover of actin in dendritic spines and its regulation by activity , 2002, Nature Neuroscience.

[22]  Wei-Yang Lu,et al.  Activation of Synaptic NMDA Receptors Induces Membrane Insertion of New AMPA Receptors and LTP in Cultured Hippocampal Neurons , 2001, Neuron.

[23]  M. Sheng,et al.  PDZ domain proteins of synapses , 2004, Nature Reviews Neuroscience.

[24]  R. Colbran,et al.  Targeting of calcium/calmodulin-dependent protein kinase II. , 2004, The Biochemical journal.

[25]  Alcino J. Silva,et al.  Genetic approaches to molecular and cellular cognition: a focus on LTP and learning and memory. , 2002, Annual review of genetics.

[26]  Ann Marie Craig,et al.  Activity Regulates the Synaptic Localization of the NMDA Receptor in Hippocampal Neurons , 1997, Neuron.

[27]  Takeharu Nagai,et al.  Rapid and persistent modulation of actin dynamics regulates postsynaptic reorganization underlying bidirectional plasticity , 2004, Nature Neuroscience.

[28]  K. Fukunaga,et al.  Immunohistochemical Localization of Ca2+/Calmodulin‐Dependent Protein Kinase II in Rat Brain and Various Tissues , 1988, Journal of neurochemistry.

[29]  R. Morris,et al.  Enhanced long-term potentiation and impaired learning in mice with mutant postsynaptic density-95 protein , 1998, Nature.

[30]  A. Craig,et al.  Role of Actin in Anchoring Postsynaptic Receptors in Cultured Hippocampal Neurons: Differential Attachment of NMDA versus AMPA Receptors , 1998, The Journal of Neuroscience.

[31]  Mary B. Kennedy,et al.  The postsynaptic density at glutamatergic synapses , 1997, Trends in Neurosciences.

[32]  A Miyawaki,et al.  Measurement of cytosolic, mitochondrial, and Golgi pH in single living cells with green fluorescent proteins. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[33]  Daniel Choquet,et al.  Differential activity-dependent regulation of the lateral mobilities of AMPA and NMDA receptors , 2004, Nature Neuroscience.

[34]  P. Greengard,et al.  Immunocytochemical localization of calcium/calmodulin-dependent protein kinase II in rat brain. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[35]  J. Lisman,et al.  The molecular basis of CaMKII function in synaptic and behavioural memory , 2002, Nature Reviews Neuroscience.

[36]  Andy Hudmon,et al.  Neuronal CA2+/calmodulin-dependent protein kinase II: the role of structure and autoregulation in cellular function. , 2002, Annual review of biochemistry.

[37]  R. Huganir,et al.  Activation of Silent Synapses by Rapid Activity-Dependent Synaptic Recruitment of AMPA Receptors , 2001, The Journal of Neuroscience.

[38]  M. Ehlers,et al.  Reinsertion or Degradation of AMPA Receptors Determined by Activity-Dependent Endocytic Sorting , 2000, Neuron.

[39]  S. Tonegawa,et al.  The Essential Role of Hippocampal CA1 NMDA Receptor–Dependent Synaptic Plasticity in Spatial Memory , 1996, Cell.

[40]  Gavin Rumbaugh,et al.  Phosphorylation of the AMPA Receptor GluR1 Subunit Is Required for Synaptic Plasticity and Retention of Spatial Memory , 2003, Cell.

[41]  K. Shen,et al.  Dynamic control of CaMKII translocation and localization in hippocampal neurons by NMDA receptor stimulation. , 1999, Science.

[42]  M. Kennedy,et al.  Signal-processing machines at the postsynaptic density. , 2000, Science.

[43]  M Neal Waxham,et al.  A Mechanism for Ca2+/Calmodulin-Dependent Protein Kinase II Clustering at Synaptic and Nonsynaptic Sites Based on Self-Association , 2005, The Journal of Neuroscience.

[44]  R. Huganir,et al.  Redistribution and Stabilization of Cell Surface Glutamate Receptors during Synapse Formation , 1997, The Journal of Neuroscience.