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Membrane Protein Identification in Rodent Brain Tissue Samples and Acute Brain Slices

1
Institute of Anatomy, University Medical Center Rostock, 18057 Rostock, Germany
2
Core Facility Proteome Analysis, University Medical Center Rostock, 18057 Rostock, Germany
*
Author to whom correspondence should be addressed.
Cells 2019, 8(5), 423; https://doi.org/10.3390/cells8050423
Received: 11 April 2019 / Revised: 7 May 2019 / Accepted: 8 May 2019 / Published: 8 May 2019
(This article belongs to the Special Issue Oligodendrocyte Physiology and Pathology Function)
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Abstract

Acute brain slices are a sample format for electrophysiology, disease modeling, and organotypic cultures. Proteome analyses based on mass spectrometric measurements are seldom used on acute slices, although they offer high-content protein analyses and explorative approaches. In neuroscience, membrane proteins are of special interest for proteome-based analysis as they are necessary for metabolic, electrical, and signaling functions, including myelin maintenance and regeneration. A previously published protocol for the enrichment of plasma membrane proteins based on aqueous two-phase polymer systems followed by mass spectrometric protein identification was adjusted to the small sample size of single acute murine slices from newborn animals and the reproducibility of the results was analyzed. For this, plasma membrane proteins of 12 acute slice samples from six animals were enriched and analyzed by liquid chromatography-mass spectrometry. A total of 1161 proteins were identified, of which 369 were assigned to membranes. Protein abundances showed high reproducibility between samples. The plasma membrane protein separation protocol can be applied to single acute slices despite the low sample size and offers a high yield of identifiable proteins. This is not only the prerequisite for proteome analysis of organotypic slice cultures but also allows for the analysis of small-sized isolated brain regions at the proteome level. View Full-Text
Keywords: plasma membrane proteins; liquid chromatography-mass spectrometry; murine acute brain slices; reproducibility; rat cerebellum plasma membrane proteins; liquid chromatography-mass spectrometry; murine acute brain slices; reproducibility; rat cerebellum
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Joost, S.; Mikkat, S.; Wille, M.; Schümann, A.; Schmitt, O. Membrane Protein Identification in Rodent Brain Tissue Samples and Acute Brain Slices. Cells 2019, 8, 423.

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