Purification and culture of nerve growth factor receptor (p75)-expressing basal forebrain cholinergic neurons

Nature Protocols
Aletta C SchnitzlerJan Krzysztof Blusztajn

Abstract

The activity of the basal forebrain cholinergic neurons (BFCNs) that innervate the cerebral cortex and hippocampus is essential for normal learning and memory. Here, we present a method to isolate and culture BFCNs from the embryonic murine septum that takes advantage of their restricted expression of the nerve growth factor receptor (p75) in conjunction with fluorescence-activated cell sorting. The septal region dissection, cell dissociation and staining process, and cell sorting parameters are described in detail. Sufficient cell yield and optimized cell culture conditions make this protocol suitable for multiple assays including immunocytochemistry, reverse transcriptase PCR, microarray profiling, acetylcholine measurements and electrophysiological assessment. The study of these neurons as a purified population will greatly advance our understanding of factors that influence their development and maintenance.

References

Sep 3, 2003·Journal of Neuroscience Methods·Solène Sergent-TanguyPhilippe Naveilhan
May 5, 2005·Proceedings of the National Academy of Sciences of the United States of America·Ignacio Lopez-CoviellaJan Krzysztof Blusztajn
Feb 3, 2006·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Timothy G J AllenDavid A Brown
Aug 31, 2006·Physiological Genomics·Yvonne N TalliniMichael I Kotlikoff
Oct 24, 2006·Nature Neuroscience·P Hande Ozdinler, Jeffrey D Macklis

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Citations

Mar 22, 2013·PloS One·Richard FairlessAdolfo Cavalié
Jun 18, 2010·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Aletta C SchnitzlerJan Krzysztof Blusztajn
Oct 2, 2012·Journal of Neuroscience Methods·Nicholas NagykeryChangiz Geula
Mar 26, 2016·Frontiers in Synaptic Neuroscience·Jan Krzysztof Blusztajn, Jasmine Rinnofner
Nov 7, 2012·Stem Cell Research·Alexander BirbrairOsvaldo Delbono
Jun 20, 2020·Translational Neurodegeneration·Ruinan ShenChengbiao Wu
Aug 30, 2020·Cellular and Molecular Neurobiology·Sarra DjemilDaniel T S Pak
Mar 3, 2015·Current Protocols in Cell Biology·Alice KlausmeyerS Wiese
Dec 5, 2009·Current Protocols in Stem Cell Biology·Jan PruszakGuido Nikkhah
Feb 5, 2021·Life Science Alliance·María Luisa FrancoMarçal Vilar

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