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Elucidating the molecular mechanisms underlying the functional diversity of synapses requires a high-resolution, sensitive, diffusion-free, quantitative localization method that allows the determination of many proteins in functionally characterized individual synapses. Array tomography permits the quantitative analysis of single synapses but has limited sensitivity, and its application to functionally characterized synapses is challenging. Here, we aim to overcome these limitations by searching the parameter space of different fixation, resin, embedding, etching, retrieval, and elution conditions. Our optimizations reveal that etching epoxy-resin-embedded ultrathin sections with Na-ethanolate and treating them with SDS dramatically increase the labeling efficiency of synaptic proteins. We also demonstrate that this method is ideal for the molecular characterization of individual synapses following paired recordings, two-photon [Ca2+] or glutamate-sensor (iGluSnFR) imaging. This method fills a missing gap in the toolbox of molecular and cellular neuroscience, helping us to reveal how molecular heterogeneity leads to diversity in function.
Pubmed ID: 32726631 RIS Download
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A national mouse monoclonal antibody generating resource for biochemical and immunohistochemical applications in mammalian brain. NeuroMabs are generated from mice immunized with synthetic and recombinant immunogens corresponding to components of the neuronal proteome as predicted from genomic and other large-scale cloning efforts. Comprehensive biochemical and immunohistochemical analyses of human, primate and non-primate mammalian brain are incorporated into the initial NeuroMab screening procedure. This yields a subset of mouse mAbs that are optimized for use in brain (i.e. NeuroMabs): for immunocytochemical-based imaging studies of protein localization in adult, developing and pathological brain samples, for biochemical analyses of subunit composition and post-translational modifications of native brain proteins, and for proteomic analyses of native brain protein networks. The NeuroMab facility was initially funded with a five-year U24 cooperative grant from NINDS and NIMH. The initial goal of the facility for this funding period is to generate a library of novel NeuroMabs against neuronal proteins, initially focusing on membrane proteins (receptors/channels/transporters), synaptic proteins, other neuronal signaling molecules, and proteins with established links to disease states. The scope of the facility was expanded with supplements from the NIH Blueprint for Neuroscience Research to include neurodevelopmental targets, the NIH Roadmap for Medical Research to include epigenetics targets, and NIH Office of Rare Diseases Research to include rare disease targets. These NeuroMabs will then be produced on a large scale and made available to the neuroscience research community on an inexpensive basis as tissue culture supernatants or purified immunoglobulin by Antibodies Inc. The UC Davis/NIH NeuroMab Facility makes NeuroMabs available directly to end users and is unable to accommodate sales to distributors for third party distribution. Note, NeuroMab antibodies are now offered through antibodiesinc.
View all literature mentionsA next-generation web-based application that aims to provide an integrated solution for both visualization and analysis of deep-sequencing data, along with simple access to public datasets.
View all literature mentionsAmerican chemical, life science and biotechnology company owned by Merck KGaA. Merger of Sigma Chemical Company and Aldrich Chemical Company. Provides organic and inorganic chemicals, building blocks, reagents, advanced materials and stable isotopes for chemical synthesis, medicinal chemistry and materials science, antibiotics, buffers, carbohydrates, enzymes, forensic tools, hematology and histology, nucleotides, proteins, peptides, amino acids and their derivatives.
View all literature mentionsSoftware suite for electrophysiology data acquisition and analysis by Molecular Devices. Used for the control and recording of voltage clamp, current clamp, and patch clamp experiments. The software suite consists of Clampex 11 Software for data acquisition, AxoScope 11 Software for background recording, Clampfit 11 Software for data analysis, and optional Clampfit Advanced Analysis Module for sophisticated and streamlined analysis.
View all literature mentionsSoftware for image processing, analysis, and editing. The software includes features such as touch capabilities, a customizable toolbar, 2D and 3D image merging, and Cloud access and options.
View all literature mentionsManufacturer and supplier of the DMZ Universal electrode pullers which produce pipettes and electrodes with a tip diameter down to 0.2 micrometers.
View all literature mentionsTHIS RESOURCE IS NO LONGER IN SERVICE. Documented on June 1,2023. Analytics platform with various sub platforms, each with specific performance capabilities for tasks such as data analysis, data management, data visualization, and data mining procedures.
View all literature mentionsThis polyclonal targets Homer 1b/c
View all literature mentionsThis polyclonal targets IgG (H+L)
View all literature mentionsThis monoclonal targets GABA-A receptor alpha1
View all literature mentionsThis polyclonal targets mouse HCN1 (NP_034538, NM_010408)
View all literature mentionsThis polyclonal targets ERC 1b/2
View all literature mentionsThis unknown targets Mouse IgG, Fc? Subclass 1 Specific
View all literature mentionsThis polyclonal targets parvalbumin
View all literature mentionsThis polyclonal targets Piccolo
View all literature mentionsThis polyclonal targets mouse GluA4, N-terminal 262-294 aa (AB022913)
View all literature mentionsThis polyclonal targets Rabbit IgG
View all literature mentionsThis unknown targets Guinea Pig IgG (H+L)
View all literature mentionsThis unknown targets Cholecystokinin, aa 26-33 (CCK-8)
View all literature mentionsThis monoclonal targets GluA1/GluR1 glutamate receptor
View all literature mentionsThis polyclonal targets Guinea pig
View all literature mentionsThis unknown targets Chicken IgY (IgG) (H+L)
View all literature mentionsThis polyclonal targets Glutamate Transporter, Glial (GLAST)
View all literature mentionsThis unknown targets Mouse IgG, Fc? Subclass 1 Specific
View all literature mentionsThis monoclonal targets Cannabinoid receptor CB1-R
View all literature mentionsThis polyclonal targets Cannabinoid receptor CB1-R
View all literature mentionsThis unknown targets ZO-1
View all literature mentionsThis polyclonal secondary targets IgG (H+L)
View all literature mentionsThis unknown targets VIP
View all literature mentionsThis polyclonal targets mouse CB1, C-terminal 31 aa (NM007726)
View all literature mentionsThis unknown targets Vesicular Glutamate Transporter 2 (VGLUT2)
View all literature mentionsThis polyclonal targets N-Cadherin
View all literature mentionsThis monoclonal targets Cav3.1 Ca2+ channel
View all literature mentionsThis unknown targets NaV1.6 (SCN8A) Channel
View all literature mentionsThis polyclonal targets VGLUT1
View all literature mentionsThis polyclonal targets Ca2+ channel (R-type, alpha-1E subunit)
View all literature mentionsThis polyclonal targets Munc 13-1
View all literature mentionsThis polyclonal targets Munc 13-1
View all literature mentionsThis polyclonal targets Ca2+ channel (P/Q-type, alpha-1A subunit)
View all literature mentionsThis polyclonal targets VGAT (cytoplasmic domain)
View all literature mentionsThis monoclonal targets Syntaxin
View all literature mentionsThis polyclonal targets mouse VAChT, 491-530 aa (AF019045)
View all literature mentionsThis polyclonal targets SynCAM 1/2/3
View all literature mentionsThis polyclonal targets mouse mGluR1a, 945-1127 aa (NM_016976)
View all literature mentionsThis polyclonal targets mouse mGluR1a, 945-1127 aa (NM_016976)
View all literature mentionsThis polyclonal targets Ca2+ channel (P/Q-type, alpha-1A subunit)
View all literature mentionsThis polyclonal targets Synaptotagmin 7
View all literature mentionsThis monoclonal targets Myelin Basic Protein (84-89)
View all literature mentionsThis polyclonal targets Synaptotagmin 2 (cytoplasmic domain)
View all literature mentionsThis monoclonal targets GFAP
View all literature mentionsThis monoclonal targets Gephyrin
View all literature mentionsThis monoclonal targets Somatostatin
View all literature mentionsThis polyclonal targets mouse M2, 296-356 aa (XM145245)
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View all literature mentionsThis polyclonal targets Synapsin I
View all literature mentionsThis monoclonal targets Calretinin (CR)
View all literature mentionsThis unknown targets SNAP-25
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View all literature mentionsThis polyclonal targets GABA-A receptor beta1
View all literature mentionsThis polyclonal targets GABA-A receptor beta2
View all literature mentionsThis monoclonal targets Kv4.3 K+ channel
View all literature mentionsThis monoclonal targets Kv2.1 K+ channel
View all literature mentionsThis polyclonal targets Cy3-Streptavidin
View all literature mentionsThis unknown targets KV4.2 Channel
View all literature mentionsThis polyclonal targets mouse Kcna2, 482-496 aa (NM_008417)
View all literature mentionsThis polyclonal targets beta-catenin
View all literature mentionsThis polyclonal targets mouse, 478-492 aa (NM_010595)
View all literature mentionsThis polyclonal targets GABA-A receptor alpha6
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View all literature mentionsThis polyclonal targets mouse, 478-492 aa (NM_010595)
View all literature mentionsThis unknown targets
View all literature mentionsThis polyclonal targets Shank 3
View all literature mentionsThis polyclonal targets Shank 3
View all literature mentionsThis polyclonal targets Bassoon
View all literature mentionsThis unknown targets Rat IgG (H+L)
View all literature mentionsThis polyclonal targets mouse HTT, 1-77 aa (AF013604)
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View all literature mentionsThis unknown targets Mouse IgG, Fc? Subclass 2b Specific
View all literature mentionsThis polyclonal targets PSD 95 (PDZ domain)
View all literature mentionsThis unknown targets Mouse IgG, Fc? Subclass 2a Specific
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View all literature mentionsThis polyclonal targets Piccolo
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View all literature mentionsThis monoclonal targets NeuN clone A60 Alexa Fluor 488 conjugated
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View all literature mentionsThis unknown targets Guinea Pig IgG (H+L)
View all literature mentionsOpen source Java based image processing software program designed for scientific multidimensional images. ImageJ has been transformed to ImageJ2 application to improve data engine to be sufficient to analyze modern datasets.
View all literature mentionsMus musculus with name B6.Cg-Gt(ROSA)26Sortm9(CAG-tdTomato)Hze/J from IMSR.
View all literature mentionsMus musculus with name STOCK Tg(Chrna2-cre)OE25Gsat/Mmucd from MMRRC.
View all literature mentions
Mus musculus with name FVB.129P2-Pde6b+ Tyr
Mus musculus with name C57BL/6J from IMSR.
View all literature mentionsRattus norvegicus with name Crl:WI from RGD.
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