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]