Protocol for a high-throughput semiautomated preparation for filtered synaptoneurosomes

Abstract

This book outlines various techniques for the isolation and application of synaptosomes for neuroscience research. One of the simplest techniques involves fractionation of synaptoneurosomes through filtration and low-speed centrifugation (Hollingsworth J Neurosci 5:2240–2253, 1985). This approach however is limited by two essential stages of the synaptoneurosome tissue preparation: the manual homogenization and filtrations are both laborious and slow. We have updated this traditional technique to include modern benchtop homogenizers and centrifugal filter units to simplify these labor-intensive stages, to make each stage faster and to reduce the variability between samples. Here we outline our protocol to produce filtered synaptoneurosomes that reduce sample preparation time, increase the amount of tissue recovered, and, most importantly, increase protein enrichment.