Proteomic screening of glutamatergic mouse brain synaptosomes isolated by fluorescence activated sorting.

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  • Additional Information
    • Source:
      Publisher: Wiley Blackwell Country of Publication: England NLM ID: 8208664 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1460-2075 (Electronic) Linking ISSN: 02614189 NLM ISO Abbreviation: EMBO J Subsets: MEDLINE
    • Publication Information:
      Publication: 2014- : London : Wiley Blackwell
      Original Publication: Eynsham, Oxford, England : Published for the European Molecular Biology Organization by IRL Press, [c1982-
    • Subject Terms:
      Brain/*cytology ; Flow Cytometry/*methods ; Glutamic Acid/*metabolism ; Neurons/*cytology ; Synaptosomes/*physiology; Animals ; Brain/metabolism ; Cell Separation/methods ; Ion Channels/metabolism ; Mice ; Mice, Knockout ; Neurons/metabolism ; Proteomics ; Synapses/metabolism ; Vesicular Glutamate Transport Protein 1/genetics ; Vesicular Glutamate Transport Protein 1/metabolism
    • Abstract:
      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.
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    • Accession Number:
      0 (FXYD6 protein, mouse)
      0 (Ion Channels)
      0 (Slc17a7 protein, mouse)
      0 (Vesicular Glutamate Transport Protein 1)
      3KX376GY7L (Glutamic Acid)
    • Publication Date:
      Date Created: 20140114 Date Completed: 20140314 Latest Revision: 20211021
    • Publication Date:
      20221213
    • Accession Number:
      PMC3989609
    • Accession Number:
      10.1002/embj.201386120
    • Accession Number:
      24413018