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

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.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0015-3761-7 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-002A-08B5-7
Genre: Journal Article

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 Creators:
Biesemann, C., Author
Groenborg, M.1, Author              
Luquet, E., Author
Wichert, S. P., Author
Bernard, V., Author
Bungers, S. R., Author
Cooper, B., Author
Varoqueaux, F., Author
Li, L., Author
Byrne, J. A., Author
Urlaub, H.2, Author              
Jahn, O., Author
Brose, N., Author
Herzog, E., Author
Affiliations:
1Department of Neurobiology, MPI for biophysical chemistry, Max Planck Society, ou_578595              
2Research Group of Bioanalytical Mass Spectrometry, MPI for biophysical chemistry, Max Planck Society, ou_578613              

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Free keywords: Fluorescence activated synaptosome sorting; proteomics; subcellular fractionation; synaptosome; vesicular glutamate transporter
 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.

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Language(s): eng - English
 Dates: 2014-01-102014-01-16
 Publication Status: Published in print
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 Rev. Method: Peer
 Identifiers: DOI: 10.1002/embj.201386120
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Title: EMBO Journal
Source Genre: Journal
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Pages: - Volume / Issue: 33 (2) Sequence Number: - Start / End Page: 157 - 170 Identifier: -