Purification of Afference-Specific Synaptosome Populations Using Fluorescence-Activated Synaptosome Sorting

Paget-Blanc V(1), Pronot M(1), Pfeffer ME(1), Angelo MF(1), Herzog E(2).

Author information:
(1)Univ. Bordeaux, CNRS, Interdisciplinary Institute for Neuroscience, IINS, UMR
5297, Bordeaux, France.
(2)Univ. Bordeaux, CNRS, Interdisciplinary Institute for Neuroscience, IINS, UMR
5297, Bordeaux, France. .

The central nervous system contains a complex intermingled network of neuronal,
glial, and vascular cells, and for several decades, neurobiologists have used
subcellular fractionation methods to analyze the molecular structure and
functional features of the different cell populations. Biochemists have
optimized fractionation protocols that enrich specific compartments such as
synapses (called « synaptosomes ») and synaptic vesicles to reduce this
complexity. However, these approaches suffered from a lack of specificity and
purity, which is why we previously extended the conventional synaptosome
preparation to purify fluorescent synaptosomes from VGLUT1venus knock-in mice on
a cell sorter. We adapted our previous protocol to sort from single neuronal
projections and small target regions of the brain as we did in the present
example by labeling dopaminergic projections to the striatum. We proved that our
newest method allows a steep enrichment in fluorescent dopaminergic synaptosomes
containing presynaptic varicosities and associated postsynaptic elements and a
substantial depletion in glial contaminants. Here we propose a detailed
procedure for implementing projection-specific fluorescence-activated
synaptosome sorting.

© 2025. The Author(s), under exclusive license to Springer Science+Business
Media, LLC, part of Springer Nature.

DOI: 10.1007/978-1-0716-4446-1_6
PMID: 40220095 [Indexed for MEDLINE]

Auteurs Bordeaux Neurocampus