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. 2023 May 12;16:1175851. doi: 10.3389/fnmol.2023.1175851

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Copyright © 2023 Gorski, Jackson, Nyman, Rezov, Battersby and Lehesjoki.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Synaptosomal fractionation and characterization. (A) Schematic presentation of synaptosome isolation from mouse cerebella (pink) using sucrose and Percoll. Synaptosome preparations consist of functional pre- and postsynaptic terminals containing organelles, synaptic vesicles, and receptors for neurotransmitters. (B) Immunoblot detection of pre- and postsynaptic proteins Synaptophysin (SYP) and Post-synaptic density 95 (PSD-95), respectively, and depletion of Histone deacetylase 2 (HDAC2) show synaptic protein enrichment in synaptosome preparations (Syn) compared to cerebellar lysate (Lys). Mitochondrial abundance is shown in Percoll-fractions by detection of Mitochondrial import receptor subunit TOM40. Cytosolic proteins are represented by β-tubulin and cyclin dependent kinase 5 (CDK5). Sucrose and Percoll-isolated fractions were used for mass spectrometry, and analyses of synaptic mitochondria, respectively. Mt., mitochondria; S, SYP; C, cytoplasm; P, PSD-95. Asterisk indicates carry-over signal.