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Acute in vivo proximity labeling for membrane targeted proteomics in neuronal circuits

ABSTRACT: The field of molecular systems neuroscience aims to bridge neuronal circuit activity patterns with changes in protein expression and localization in the context of specific animal behaviors. However, there are limited tools for capturing changes in subcellulary-defined proteomes within neuronal circuits during activity-gated timescales in vivo. Here, we engineered targeted versions of the proximity labeling enzyme TurboID, to tag proteins at the neuronal membrane during a user-delivered biotin injection. We optimized a labeling strategy that enables a one-to-two-hour labeling window to tag proteins in prefrontal cortex (PFC) cell bodies and corresponding axons in two downstream projections. We performed proteomics to identify proteins enriched across different projections, and identified membrane associated proteins in the PFC cell bodies that are enriched at the membrane following an acute cocaine injection. These advancements enable the detection of proteins at the subcellular level within short labeling windows, allowing identification of stimulus-specific proteomes in behaving mice.

INSTRUMENT(S): timsTOF HT, Orbitrap Exploris 480

ORGANISM(S): Mus Musculus (ncbitaxon:10090)

SUBMITTER: Christina K. Kim1

PROVIDER: MSV000099974 | MassIVE | Thu Nov 20 12:00:00 GMT 2025

SECONDARY ACCESSION(S): PXD071005

REPOSITORIES: MassIVE

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