Synaptoneurosome micromethod for fractionation of mouse and human brain, and primary neuronal cultures

[Display omitted] ► Easy step-by-step protocol for preparing synaptoneurosomes (SNs) from small amounts of mammalian brain tissue. ► Novel SN applications including resected human neocortex and rat primary cortical neuronal cultures. ► New tool for synaptic biology including SNs prepared from human neurodegenerative diseases for example AD and PD. Brain and primary neuron fractions enriched in synaptic terminals are important tools for neuroscientists in biochemical, neuroanatomical and physiological studies. We describe an annotated updated micro-method for preparing synaptoneurosomes (SNs) enriched in presynaptic and postsynaptic elements. An easy to follow, step-by-step, protocol is provided for making SNs from small amounts of mammalian brain tissue. This includes novel applications for material obtained from human neurosurgical procedures and primary rat neuronal cultures. Our updated method for preparing SNs using smaller amounts of tissue provides a valuable new tool and expands the capabilities of neuroscientists.