Proteomic screening of glutamatergic mouse brain synaptosomes isolated by fluorescence activated sorting.
EMBO J. 2014-01-01; 33(2): 157-170
Lire sur PubMed
1. EMBO J. 2014 Jan 13;33(2):157-70. doi: 10.1002/embj.201386120. Epub 2014 Jan 10.
Proteomic screening of glutamatergic mouse brain synaptosomes isolated by
fluorescence activated sorting.
Biesemann C(1), Grønborg M, Luquet E, Wichert SP, Bernard V, Bungers SR, Cooper
B, Varoqueaux F, Li L, Byrne JA, Urlaub H, Jahn O, Brose N, Herzog E.
(1)Department of Molecular Neurobiology, Max Planck Institute of Experimental
Medicine, Göttingen, Germany.
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
PMID: 24413018 [Indexed for MEDLINE]