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Journal Article

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

MPS-Authors
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Groenborg,  M.
Department of Neurobiology, MPI for biophysical chemistry, Max Planck Society;

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Urlaub,  H.
Research Group of Bioanalytical Mass Spectrometry, MPI for biophysical chemistry, Max Planck Society;

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Fulltext (public)

1921868.pdf
(Publisher version), 4MB

Supplementary Material (public)

1921868_Suppl_1.htm
(Supplementary material), 394KB

Citation

Biesemann, C., Groenborg, M., Luquet, E., Wichert, S. P., Bernard, V., Bungers, S. R., et al. (2014). Proteomic screening of glutamatergic mouse brain synaptosomes isolated by fluorescence activated sorting. EMBO Journal, 33(2), 157-170. doi:10.1002/embj.201386120.


Cite as: http://hdl.handle.net/11858/00-001M-0000-0015-3761-7
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.