Preparation of Synaptoneurosomes to Study the Synapse in the Murine Cerebral Cortex

• Published in: Bio-protocol Vol. 11; no. 2; p. e3896
• Main Authors: Diaz, Ariel, Torre, Enrique, Yepes, Manuel
• Format: Journal Article
• Language: English
• Published: United States Bio-Protocol 20-01-2021; Bio-protocol LLC
• Subjects: Biology; Methods; Brain; Subcellular fractionation; Synapse; Synaptoneurosomes; Synaptic compartment; Tripartite synapse
• ISSN: 2331-8325
• DOI: 10.21769/BioProtoc.3896

The synapse is a complex structure where the transmission of information takes place. Synaptic dysfunction is one of the earliest pathophysiological events in several diseases, such as traumatic brain injury, cerebral ischemia, and neurodegenerative diseases. Thus, a methodology to study synaptic structure and function is crucial for the development of potential strategies for the treatment of many neurological diseases. Synaptoneurosomes (SNs) are structures assembled by the sealed presynaptic bouton and the attached post-synaptic density. Despite the fact that for a long time it has been recognized that SNs are a powerful tool to study synaptic function, composition, and structure, its use has been limited by the requirement of relatively large amounts of material to successfully isolate them. Here we describe a three-step centrifugation procedure performed under hypotonic conditions to isolate SNs from small volumes of the cerebral cortex.