title: Purification of Synaptosome Populations Using Fluorescence-Activated Synaptosome Sorting.
abstract: For several decades, neurobiologists have used subcellular fractionation methods to analyze the molecular structure and some functional features of the cells in the central nervous system. Indeed, brain tissue contains a complex intermingled network of neuronal, glial, and vascular cells. To reduce this complexity biochemists have optimized fractionation protocols that enrich in specific compartments such as synapses (called "synaptosomes") and synaptic vesicles, for example. However, recently, these approaches suffered from a lack of specificity and purity. In a recent effort, we extended the conventional synaptosome preparation to purify fluorescent synaptosomes on a cell sorter. We could prove that our method allows for the steep enrichment in fluorescent excitatory VGLUT1venussynaptosomes containing the presynaptic element and the tip of the post-synaptic element and a strong depletion in neuronal and glial contaminants. Here, we propose a detailed procedure for the implementation of Fluorescence Activated Synaptosome Sorting.
authors.affiliation: Interdisciplinary Institute for Neuroscience, University Bordeaux, CNRS, UMR 5297, F-33000, Bordeaux, France.|Interdisciplinary Institute for NeuroScience, CNRS, UMR 5297, F-33000, Bordeaux, France.
authors.forename: Elisa
authors.initials: E
authors.lastname: Luquet
authors.affiliation: Department of Molecular Neurobiology, Max Planck Institute of Experimental Medicine, 37075, Göttingen, Germany.
authors.forename: Christoph
authors.initials: C
authors.lastname: Biesemann
authors.affiliation: Institut de Biologie Paris Seine, University Pierre et Marie Curie, IBPS, F-75005, Paris, France.
authors.forename: Annie
authors.initials: A
authors.lastname: Munier
authors.affiliation: Interdisciplinary Institute for Neuroscience, University Bordeaux, CNRS, UMR 5297, F-33000, Bordeaux, France. etienne.herzog@u-bordeaux2.fr.|Interdisciplinary Institute for NeuroScience, CNRS, UMR 5297, F-33000, Bordeaux, France. etienne.herzog@u-bordeaux2.fr.
authors.forename: Etienne
authors.initials: E
authors.lastname: Herzog
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism; D002458:Cell Fractionation / Q000379:methods*; D005434:Flow Cytometry / Q000379:methods*; D051379:Mice; D013569:Synapses / Q000378:metabolism; D009435:Synaptic Transmission; D013572:Synaptic Vesicles / Q000378:metabolism; D013574:Synaptosomes / Q000378:metabolism*; D014461:Ultracentrifugation / Q000379:methods
keywords: Cytometry; Fluorescence activated synaptosome sorting; Glutamatergic neurotransmission; Micro-particle sorting; Subcellular fractionation; Synapse; Synaptosome; VGLUT1