citation_mjid: embojnl;33/2/157
article:published_time: 2014-01-13
citation_title: Proteomic screening of glutamatergic mouse brain synaptosomes isolated by fluorescence activated sorting
citation_author_institution: Max Planck Institute of Experimental Medicine; Max Planck Institute of Biophysical Chemistry; CNRS; Max Planck Institute of Experimental Medicine; INSERM U952; Max Planck Institute of Experimental Medicine; Max Planck Institute of Experimental Medicine; Max Planck Institute of Experimental Medicine; Max Planck Institute of Experimental Medicine; Kids Research Institute; Max Planck Institute for Biophysical Chemistry; Max Planck Institute of Experimental Medicine; Max Planck Institute of Experimental Medicine; Max Planck Institute of Experimental Medicine
og:site_name: EMBO J
keywords: Fluorescence Activated Synaptosome Sorting, proteomics, subcellular fractionation, synaptosome, vesicular glutamate transporter
citation_reference: citation_journal_title=EMBO Journal;citation_pages=157-170;citation_volume=33;citation_year=2014
citation_publisher: EMBO Press
citation_journal_title: The EMBO Journal
citation_id: 33/2/157
citation_date: 01/13/2014
title: Proteomic screening of glutamatergic mouse brain synaptosomes isolated by fluorescence activated sorting | EMBO J
DC.Description: For decades, neuroscientists have used enriched preparations of synaptic particles called synaptosomes to study synapse function. However, the interpretation of corresponding data is problematic as synaptosome preparations contain multiple types of synapses and non?synaptic neuronal and glial contaminants. We established a novel Fluorescence Activated Synaptosome Sorting (FASS) method that substantially improves conventional synaptosome enrichment protocols and enables high?resolution biochemical analyses of specific synapse subpopulations. Employing knock?in mice with fluorescent glutamatergic synapses, we show that FASS isolates intact ultrapure synaptosomes composed of a resealed presynaptic terminal and a postsynaptic density as assessed by light and electron microscopy. FASS synaptosomes contain bona fide glutamatergic synapse proteins but are almost devoid of other synapse types and extrasynaptic or glial contaminants. We identified 163 enriched proteins in FASS samples, of which FXYD6 and Tpd52 were validated as new synaptic proteins. FASS purification thus enables high?resolution biochemical analyses of specific synapse subpopulations in health and disease.  ![][1]</img>  This study describes the establishment, validation, and application of a new fluorescence?activated sorting method ? Fluorescence Activated Synaptosome Sorting or FASS ? that allows for purification of glutamatergic synaptosomes from knock?in mutant mice expressing the fluorescent synaptic marker protein VGLUT1?Venus.   EMBO Journal (2014) 33, 157?170   [1]: pending:yes
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dc:title: Proteomic screening of glutamatergic mouse brain synaptosomes isolated by fluorescence activated sorting | EMBO J
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DC.Publisher: EMBO Press
citation_pdf_url: http://emboj.embopress.org/content/33/2/157.full.pdf
citation_section: Resource
DC.Contributor: Christoph Biesemann
citation_lastpage: 170
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article:section: Resource
citation_pmid: 24413018
DC.Identifier: 10.1002/embj.201386120
DC.Rights: © 2013 The Authors
og:title: Proteomic screening of glutamatergic mouse brain synaptosomes isolated by fluorescence activated sorting
DC.Title: Proteomic screening of glutamatergic mouse brain synaptosomes isolated by fluorescence activated sorting
citation_abstract_html_url: http://emboj.embopress.org/content/33/2/157.abstract
citation_issue: 2
citation_authors: Biesemann, Christoph; Grønborg, Mads; Luquet, Elisa; Wichert, Sven P; Bernard, Véronique; Bungers, Simon R; Cooper, Ben; Varoqueaux, Frédérique; Li, Liyi; Byrne, Jennifer A; Urlaub, Henning; Jahn, Olaf; Brose, Nils; Herzog, Etienne
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citation_firstpage: 157
DC.Language: en
citation_doi: 10.1002/embj.201386120
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DC.Date: 2014-01-13
citation_volume: 33
og:url: http://emboj.embopress.org/content/33/2/157
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