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

doi:10.1016/j.xpro.2023.102061

Fast and efficient synaptosome isolation and post-synaptic density enrichment from hiPSC-motor neurons by biochemical sub-cellular fractionation

STAR Protoc. 2023 Mar 17;4(1):102061. doi: 10.1016/j.xpro.2023.102061. Epub 2023 Jan 25.

Authors

Sandeep Rajkumar¹, Tobias M Böckers², Alberto Catanese³

Affiliations

¹ Institute for Anatomy and Cell Biology, Ulm University, 89081 Ulm, Germany. Electronic address: sandeep.rajkumar@live.com.
² Institute for Anatomy and Cell Biology, Ulm University, 89081 Ulm, Germany; German Center for Neurodegenerative Diseases (DZNE), Ulm Site, 89081 Ulm, Germany.
³ Institute for Anatomy and Cell Biology, Ulm University, 89081 Ulm, Germany; German Center for Neurodegenerative Diseases (DZNE), Ulm Site, 89081 Ulm, Germany. Electronic address: alberto.catanese@uni-ulm.de.

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.

Keywords: Cell culture; Cell separation/fractionation; Neuroscience; Stem Cells.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Induced Pluripotent Stem Cells*
Motor Neurons
Post-Synaptic Density
Synaptosomes* / metabolism