Isolation of Neuronal Synaptic Membranes by Sucrose Gradient Centrifugation.

Autor:	Hopiavuori BR; Oklahoma Center for Neuroscience, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA.; Department of Ophthalmology, Dean McGee Eye Institute, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA., Masser DR; Department of Physiology, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA.; Reynolds Oklahoma Center on Aging, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA., Wilkerson JL; Department of Ophthalmology, Dean McGee Eye Institute, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA.; Department of Cell Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA., Brush RS; Department of Ophthalmology, Dean McGee Eye Institute, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA., Mandal NA; Oklahoma Center for Neuroscience, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA.; Department of Ophthalmology, Dean McGee Eye Institute, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA.; Department of Physiology, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA.; Department of Cell Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA., Anderson RE; Oklahoma Center for Neuroscience, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA.; Department of Ophthalmology, Dean McGee Eye Institute, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA.; Department of Cell Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA., Freeman WM; Genes & Human Disease Program, Oklahoma Medical Research Foundation, Oklahoma City, USA. bill-freeman@omrf.org.
Jazyk:	angličtina
Zdroj:	Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2023; Vol. 2625, pp. 7-15.
DOI:	10.1007/978-1-0716-2966-6_2
Abstrakt:	Sucrose gradient centrifugation is a very useful technique for isolating specific membrane types based on their size and density. This is especially useful for detecting fatty acids and lipid molecules that are targeted to specialized membranes. Without fractionation, these types of molecules could be below the levels of detection after being diluted out by the more abundant lipid molecules with a more ubiquitous distribution throughout the various cell membranes. Isolation of specific membrane types where these lipids are concentrated allows for their detection and analysis. We describe herein our synaptic membrane isolation protocol that produces excellent yield and clear resolution of five major membrane fractions from a starting neural tissue homogenate: P1 (nuclear), P2 (cytoskeletal), P3 (neurosynaptosomal), PSD (post-synaptic densities), and SV (synaptic vesicle). (© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)
Databáze:	MEDLINE
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