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Titel: Enrichment and isolation of neurons from adult mouse brain for ex vivo analysis
VerfasserIn: Berl, Sabina
Karram, Khalad
Scheller, Anja
Jungblut, Melanie
Kirchhoff, Frank
Waisman, Ari
Sprache: Englisch
Titel: Journal of Neuroscience Methods
Bandnummer: 283
Seiten: 15-22
Verlag/Plattform: Elsevier
Erscheinungsjahr: 2017
DDC-Sachgruppe: 610 Medizin, Gesundheit
Dokumenttyp: Journalartikel / Zeitschriftenartikel
Abstract: Background Isolation of neurons from the adult mouse CNS is important in order to study their gene expression during development or the course of different diseases. New methods Here we present two different methods for the enrichment or isolation of neurons from adult mouse CNS. These methods: are either based on flow cytometry sorting of eYFP expressing neurons, or by depletion of non-neuronal cells by sorting with magnetic-beads. Results Enrichment by FACS sorting of eYFP positive neurons results in a population of 62.4% NeuN positive living neurons. qPCR data shows a 3–5fold upregulation of neuronal markers. The isolation of neurons based on depletion of non-neuronal cells using the Miltenyi Neuron Isolation Kit, reaches a purity of up to 86.5%. qPCR data of these isolated neurons shows an increase in neuronal markers and an absence of glial markers, proving pure neuronal RNA isolation. Comparison with existing methods Former data related to neuronal gene expression are mainly based on histology, which does not allow for high-throughput transcriptome analysis to examine differential gene expression. Conclusion These protocols can be used to study cell type specific gene expression of neurons to unravel their function in the process of damage to the CNS.
DOI der Erstveröffentlichung: 10.1016/j.jneumeth.2017.03.015
URL der Erstveröffentlichung: https://www.sciencedirect.com/science/article/abs/pii/S0165027017300717
Link zu diesem Datensatz: urn:nbn:de:bsz:291--ds-395699
hdl:20.500.11880/35664
http://dx.doi.org/10.22028/D291-39569
ISSN: 0165-0270
Datum des Eintrags: 18-Apr-2023
Fakultät: M - Medizinische Fakultät
Fachrichtung: M - Physiologie
Professur: M - Prof. Dr. Frank Kirchhoff
Sammlung:SciDok - Der Wissenschaftsserver der Universität des Saarlandes

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