title: Preparation of synaptic plasma membrane and postsynaptic density proteins using a discontinuous sucrose gradient.
abstract: Neuronal subcellular fractionation techniques allow the quantification of proteins that are trafficked to and from the synapse. As originally described in the late 1960's, proteins associated with the synaptic plasma membrane can be isolated by ultracentrifugation on a sucrose density gradient. Once synaptic membranes are isolated, the macromolecular complex known as the post-synaptic density can be subsequently isolated due to its detergent insolubility. The techniques used to isolate synaptic plasma membranes and post-synaptic density proteins remain essentially the same after 40 years, and are widely used in current neuroscience research. This article details the fractionation of proteins associated with the synaptic plasma membrane and post-synaptic density using a discontinuous sucrose gradient. Resulting protein preparations are suitable for western blotting or 2D DIGE analysis.
authors.affiliation: Department of Pharmacology and Toxicology, University of Toronto.
authors.forename: Marie Kristel
authors.initials: MK
authors.lastname: Bermejo
authors.affiliation: Department of Pharmacology and Toxicology, University of Toronto.
authors.forename: Marija
authors.initials: M
authors.lastname: Milenkovic
authors.affiliation: Department of Pharmacology and Toxicology, University of Toronto.
authors.forename: Ali
authors.initials: A
authors.lastname: Salahpour
authors.affiliation: Department of Pharmacology and Toxicology, University of Toronto; a.ramsey@utoronto.ca.
authors.forename: Amy J
authors.initials: AJ
authors.lastname: Ramsey
mesh_terms: D000818:Animals; D002499:Centrifugation, Density Gradient / Q000379:methods*; D008565:Membrane Proteins / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism; D051379:Mice; D009419:Nerve Tissue Proteins / Q000737:chemistry* / Q000378:metabolism; D013395:Sucrose / Q000737:chemistry; D013570:Synaptic Membranes / Q000737:chemistry* / Q000378:metabolism; D013574:Synaptosomes / Q000737:chemistry
chemical_list: D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; C033652:postsynaptic density proteins; D013395:Sucrose