Purification of Synaptosome Populations Using Fluorescence-Activated Synaptosome Sorting
Methods in Molecular Biology. 2016-12-11; : 121-134
DOI: 10.1007/978-1-4939-6688-2_10
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1. Methods Mol Biol. 2017;1538:121-134.
Purification of Synaptosome Populations Using Fluorescence-Activated Synaptosome
Sorting.
Luquet E(1)(2), Biesemann C(3), Munier A(4), Herzog E(5)(6).
Author information:
(1)Interdisciplinary Institute for Neuroscience, University Bordeaux, CNRS, UMR
5297, F-33000, Bordeaux, France.
(2)Interdisciplinary Institute for NeuroScience, CNRS, UMR 5297, F-33000,
Bordeaux, France.
(3)Department of Molecular Neurobiology, Max Planck Institute of Experimental
Medicine, 37075, Göttingen, Germany.
(4)Institut de Biologie Paris Seine, University Pierre et Marie Curie, IBPS,
F-75005, Paris, France.
(5)Interdisciplinary Institute for Neuroscience, University Bordeaux, CNRS, UMR
5297, F-33000, Bordeaux, France. .
(6)Interdisciplinary Institute for NeuroScience, CNRS, UMR 5297, F-33000,
Bordeaux, France. .
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
VGLUT1venus synaptosomes 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.
DOI: 10.1007/978-1-4939-6688-2_10
PMID: 27943188 [Indexed for MEDLINE]