Preparing Synaptoneurosomes from Adult Mouse Forebrain

Many neuroscience studies involve subcellular fractionation to produce isolated or enriched synaptic fractions. Synaptosomes are prepared by flotation of synaptic membranes on sucrose or Percoll gradients. Alternatively, synaptoneurosomes are prepared by filtration of tissue homogenate through a series of filters to obtain a fraction that is enriched in pinched-off dendritic spines. Whereas the protocol for making synaptosomes is reasonably well standardized and well described in the literature, there is (to our knowledge) no detailed lab protocol for making synaptoneurosomes. Here, we give the methods used in our laboratory to produce synaptoneurosomes that are suitable for studying RNAs and proteins.