Isolation of Neuronal Synaptic Membranes by Sucrose Gradient Centrifugation

• DOI: 10.1007/978-1-0716-2966-6_2
• Publication Date: 2023-01-18T17:03:06Z
• Authors: Blake R. Hopiavuori; Dustin R. Masser; Joseph L. Wilkerson; Richard S. Brush; Nawajes A. Mandal; Robert E. Anderson; Willard M. Freeman
• Keywords: Sucrose gradient; Density gradient; Synaptic membrane; Differential centrifugation; Cell fractionation

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).