Fast and efficient synaptosome isolation and post-synaptic density enrichment from hiPSC-motor neurons by biochemical sub-cellular fractionation
Sandeep Rajkumar, Tobias M. Böckers, Alberto Catanese
Abstract
We describe here a time-efficient, in-house protocol for synaptosome isolation and enrichment of the post-synaptic density (PSD) from hiPSC-derived motor neurons. By using biochemical sub-cellular fractionation, the crude synaptosome is first isolated from the cytosol and is then further separated into the synaptic cytosol and the enriched PSD fraction. The protocol can also potentially be adapted to other hiPSC-derived neuronal types, with necessary changes made to cell seeding density and buffer volumes.
Topics & Concepts
SynaptosomeFractionationCell fractionationCytosolChemistryBiophysicsBiologyNeuroscienceBiochemistryChromatographyCentral nervous systemEnzymeNeuroscience and Neuropharmacology ResearchCellular transport and secretionMitochondrial Function and Pathology