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Preparation of Synaptoneurosomes for the Study of Glutamate Receptor Function

  • Cara J. WestmarkEmail author
  • Pamela R. Westmark
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1941)

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.

Key words

Amyloid precursor protein (APP) Fragile X mental retardation protein (FMRP) Gradient Metabotropic glutamate receptor (mGluR) Percoll Postsynaptic density Protein synthesis Synaptoneurosomes 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of NeurologyUniversity of WisconsinMadisonUSA

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