title: A high-throughput semi-automated preparation for filtered synaptoneurosomes.
abstract: BACKGROUND
  Synaptoneurosomes have become an important tool for studying synaptic proteins. The filtered synaptoneurosomes preparation originally developed by Hollingsworth et al. (1985) is widely used and is an easy method to prepare synaptoneurosomes. The hand processing steps in that preparation, however, are labor intensive and have become a bottleneck for current proteomic studies using synaptoneurosomes. For this reason, we developed new steps for tissue homogenization and filtration that transform the preparation of synaptoneurosomes to a high-throughput, semi-automated process.
  
  
  METHODS
  We implemented a standardized protocol with easy to follow steps for homogenizing multiple samples simultaneously using a FastPrep tissue homogenizer (MP Biomedicals, LLC) and then filtering all of the samples in centrifugal filter units (EMD Millipore, Corp).
  
  
  RESULTS
  The new steps dramatically reduce the time to prepare synaptoneurosomes from hours to minutes, increase sample recovery, and nearly double enrichment for synaptic proteins. These steps are also compatible with biosafety requirements for working with pathogen infected brain tissue.
  
  
  CONCLUSIONS
  The new high-throughput semi-automated steps to prepare synaptoneurosomes are timely technical advances for studies of low abundance synaptic proteins in valuable tissue samples.
authors.affiliation: McMaster Integrative Neuroscience Discovery and Study Program, McMaster University, Canada; Psychology, Neuroscience & Behavior, McMaster University, Canada. Electronic address: kmurphy@mcmaster.ca.
authors.forename: Kathryn M
authors.initials: KM
authors.lastname: Murphy
authors.affiliation: McMaster Integrative Neuroscience Discovery and Study Program, McMaster University, Canada.
authors.forename: Justin
authors.initials: J
authors.lastname: Balsor
authors.affiliation: McMaster Integrative Neuroscience Discovery and Study Program, McMaster University, Canada.
authors.forename: Simon
authors.initials: S
authors.lastname: Beshara
authors.affiliation: McMaster Integrative Neuroscience Discovery and Study Program, McMaster University, Canada.
authors.forename: Caitlin
authors.initials: C
authors.lastname: Siu
authors.affiliation: McMaster Integrative Neuroscience Discovery and Study Program, McMaster University, Canada.
authors.forename: Joshua G A
authors.initials: JG
authors.lastname: Pinto
mesh_terms: D000818:Animals; D001331:Automation; D002415:Cats; D002458:Cell Fractionation / Q000379:methods*; D002498:Centrifugation / Q000379:methods; D005374:Filtration / Q000379:methods*; D005625:Frontal Lobe / Q000737:chemistry; D015151:Immunoblotting; D009419:Nerve Tissue Proteins / Q000032:analysis; D020318:Rats, Long-Evans; D013003:Somatosensory Cortex / Q000737:chemistry / Q000648:ultrastructure; D013574:Synaptosomes / Q000737:chemistry; D013997:Time Factors
chemical_list: D009419:Nerve Tissue Proteins
keywords: Centrifugal filtration; High-throughput; Synaptic proteins; Synaptoneurosomes