SAP97 directs NMDA receptor spine targeting and synaptic plasticity

Liu, Dong; Specht, Christian G.; Waites, Clarissa L.; Butler-Munro, Charlotte; Leal-Ortiz, Sergio; Foote, Janie W.; Genoux, David; Garner, Craig C.; Montgomery, Johanna M.

doi:10.1113/jphysiol.2011.215566

Cited by 42 publications

(69 citation statements)

References 55 publications

(113 reference statements)

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“…The most obvious exception is Dlg1/Sap97, for which we did not observe activitydependent redistribution. A similar flat Dlg1 profile has been observed for different stimulation protocols in primary neuronal cultures at comparable time points (31).…”

Section: Discussionsupporting

confidence: 73%

Activity-dependent Protein Dynamics Define Interconnected Cores of Co-regulated Postsynaptic Proteins

Trinidad

Thalhammer²,

Burlingame

et al. 2013

Molecular & Cellular Proteomics

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Synapses are highly dynamic structures that mediate cellcell communication in the central nervous system. Their molecular composition is altered in an activity-dependent fashion, which modulates the efficacy of subsequent synaptic transmission events. Whereas activity-dependent trafficking of individual key synaptic proteins into and out of the synapse has been characterized previously, global activity-dependent changes in the synaptic proteome have not been studied.To test the feasibility of carrying out an unbiased largescale approach, we investigated alterations in the molecular composition of synaptic spines following mass stimulation of the central nervous system induced by pilocarpine. We observed widespread changes in relative synaptic abundances encompassing essentially all proteins, supporting the view that the molecular composition of the postsynaptic density is tightly regulated. In most cases, we observed that members of gene families displayed coordinate regulation even when they were not known to physically interact. Excitatory synaptic transmission is the primary mode of cell-cell communication in the central nervous system. The efficacy of synaptic transmission is highly regulated, and alterations in the strength of synaptic signaling within networks of neurons provide a mechanism for learning and memory storage, as well as for overall network stability. Modulation of synapse efficacy can occur through alterations in the structure and composition of the postsynaptic spine. The synaptic abundance of several molecules has been shown to be regulated in response to activity (1).The levels of individual proteins at postsynaptic spines are regulated through multiple processes. Active transport mechanisms exist and have been well characterized for AMPA-type glutamate receptors (AMPA-Rs) 1 via either insertion into the synapse or tighter association with the postsynaptic density (PSD) following lateral diffusion within the cell membrane (2). In addition to AMPA-Rs, other proteins known to be subject to activity-dependent regulation include calcium calmodulin-dependent protein kinase II alpha and beta, NMDA-type glutamate receptors (NMDA-Rs), and proteosome subunits (3-5). Synaptic protein content is dysregulated in a number of neuropsychiatric and neurodegenerative diseases, including Alzheimer's disease and fragile X mental retardation (6 -8).Most studies reported thus far have focused on a small number of selected molecules in individual experiments using a subset of synapses. Whereas learning and memory rely on the differential response of individual synapses to their specific input patterns, overall network excitability has to be maintained by homeostatic means. This homeostasis is governed by multiple pathways, and very little is known about the principles that regulate synaptic protein content across large numbers of synapses and neurons. The contributions of individual pathways and the interactions among them are largely unknown.In order to explore synaptic dynamics with a global view, we took adv...

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Section: Discussionsupporting

confidence: 73%

Activity-dependent Protein Dynamics Define Interconnected Cores of Co-regulated Postsynaptic Proteins

Trinidad

Thalhammer²,

Burlingame

et al. 2013

Molecular & Cellular Proteomics

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“…Paired whole-cell patch-clamp recordings to measure synaptic transmission between pyramidal neurons in vitro were performed as described previously (Waites et al, 2009;Li et al, 2011). Briefly, hippocampal neurons from male and female P0 Wistar rat pups were plated on 12 mm round glass coverslips, transferred to a recording chamber mounted on an Olympus BX-51 microscope, and perfused at room temperature with artificial CSF (ACSF) (in mM): 119 NaCl, 2.5 KCl, 1.3 MgSO 4 , 2.5 CaCl 2 , 1 Na 2 HPO 4 , 26.2 NaHCO 3 , and 11 glucose).…”

Section: Methodsmentioning

confidence: 99%

Autism-Associated Mutations in ProSAP2/Shank3 Impair Synaptic Transmission and Neurexin–Neuroligin-Mediated Transsynaptic Signaling

et al. 2012

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Mutations in several postsynaptic proteins have recently been implicated in the molecular pathogenesis of autism and autism spectrum disorders (ASDs), including Neuroligins, Neurexins, and members of the ProSAP/Shank family, thereby suggesting that these genetic forms of autism may share common synaptic mechanisms. Initial studies of ASD-associated mutations in ProSAP2/Shank3 support a role for this protein in glutamate receptor function and spine morphology, but these synaptic phenotypes are not universally penetrant, indicating that other core facets of ProSAP2/Shank3 function must underlie synaptic deficits in patients with ASDs. In the present study, we have examined whether the ability of ProSAP2/Shank3 to interact with the cytoplasmic tail of Neuroligins functions to coordinate pre/postsynaptic signaling through the Neurexin-Neuroligin signaling complex in hippocampal neurons of Rattus norvegicus. Indeed, we find that synaptic levels of ProSAP2/Shank3 regulate AMPA and NMDA receptor-mediated synaptic transmission and induce widespread changes in the levels of presynaptic and postsynaptic proteins via Neurexin-Neuroligin transsynaptic signaling. ASD-associated mutations in ProSAP2/Shank3 disrupt not only postsynaptic AMPA and NMDA receptor signaling but also interfere with the ability of ProSAP2/Shank3 to signal across the synapse to alter presynaptic structure and function. These data indicate that ASD-associated mutations in a subset of synaptic proteins may target core cellular pathways that coordinate the functional matching and maturation of excitatory synapses in the CNS.

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“…Whilst the same electrode solution is used for postsynaptic neurons, cesium gluconate (120 mM) is typically used as the major salt in the postsynaptic electrode, plus 5 mM QX314. NOTE: This enables stable voltage clamping at positive potentials to record postsynaptic NMDAR-mediated currents [8][9][10]13 . It also prevents the potassium-induced hyperexcitability of the presynaptic neuron by the internal solution flowing from the postsynaptic recording electrode before a giga ohm seal is made.…”

Section: Paired Whole Cell Recordingsmentioning

confidence: 99%

“…The use of organotypic brain slice cultures circumvents this obstacle as synaptic connectivity can re-establish in vitro and moreover the nature of the resulting connectivity is similar to that in native brain tissue 20 . In addition, organotypic cultures express LTP, LTD [7][8][9][10][12][13][14][15]21 and additional forms of short-term synaptic plasticity including pairedpulse facilitation (PPF) and depression (PPD) 6,22,23 , enabling plasticity mechanisms to be studied in pairs of neurons. Here we describe the detailed methodology involved in successfully attaining paired recordings in this in vitro system.…”

Section: Introductionmentioning

confidence: 99%

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Paired Whole Cell Recordings in Organotypic Hippocampal Slices

Fourie

Király

Madison

et al. 2014

JoVE

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Pair recordings involve simultaneous whole cell patch clamp recordings from two synaptically connected neurons, enabling not only direct electrophysiological characterization of the synaptic connections between individual neurons, but also pharmacological manipulation of either the presynaptic or the postsynaptic neuron. When carried out in organotypic hippocampal slice cultures, the probability that two neurons are synaptically connected is significantly increased. This preparation readily enables identification of cell types, and the neurons maintain their morphology and properties of synaptic function similar to that in native brain tissue. A major advantage of paired whole cell recordings is the highly precise information it can provide on the properties of synaptic transmission and plasticity that are not possible with other more crude techniques utilizing extracellular axonal stimulation. Paired whole cell recordings are often perceived as too challenging to perform. While there are challenging aspects to this technique, paired recordings can be performed by anyone trained in whole cell patch clamping provided specific hardware and methodological criteria are followed. The probability of attaining synaptically connected paired recordings significantly increases with healthy organotypic slices and stable micromanipulation allowing independent attainment of pre- and postsynaptic whole cell recordings. While CA3-CA3 pyramidal cell pairs are most widely used in the organotypic slice hippocampal preparation, this technique has also been successful in CA3-CA1 pairs and can be adapted to any neurons that are synaptically connected in the same slice preparation. In this manuscript we provide the detailed methodology and requirements for establishing this technique in any laboratory equipped for electrophysiology.

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SAP97 directs NMDA receptor spine targeting and synaptic plasticity

Cited by 42 publications

References 55 publications

Activity-dependent Protein Dynamics Define Interconnected Cores of Co-regulated Postsynaptic Proteins

Activity-dependent Protein Dynamics Define Interconnected Cores of Co-regulated Postsynaptic Proteins

Autism-Associated Mutations in ProSAP2/Shank3 Impair Synaptic Transmission and Neurexin–Neuroligin-Mediated Transsynaptic Signaling

Paired Whole Cell Recordings in Organotypic Hippocampal Slices

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