Preparation of Synaptoneurosomes for the Study of Glutamate Receptor Function.

Abstract

The use of synaptoneurosomes (SN) enables the detection of synaptic activity including the assessment of glutamate receptor function. SN are normally prepared by filtration and centrifugation methods. Here we review the preparation of SN by Percoll density gradient methodology for downstream applications that assesses glutamate receptor function such as measuring de novo protein synthesis. Major procedural steps include preparation of discontinuous Percoll-sucrose density gradients, collection of brain tissue, preparation of brain homogenates, isolation of synaptoneurosome bands from the discontinuous Percoll-sucrose gradients, and radiolabeling SN proteins. De novo protein synthesis can be reproducibly measured in SN prepared by this method.