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Patch-Clamp Proteomics of Single Neurons in Tissue Using Electrophysiology and Subcellular Capillary Electrophoresis Mass Spectrometry

ABSTRACT: In this project, we enabled mass spectrometry based proteomics characterization of single neurons in sections of the mouse brain. We accessed cells in a whole-cell patch clamp configuration and aspirated a portion of the neuronal soma into the patch clamp probe. The collected aspirate was expelled into a microvial, where proteins were extracted and processed for shot-gun analysis. The resulting trace amounts of protein digests were analyzed on a custom-built microanalytical capillary electrophoresis platform that was connected to an electrospray ionization high-resolution mass spectrometer. This “patch proteomics” technology identified ~150 proteins from triplicate measurement of single dopaminergic neurons in the mouse substantia nigra.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Brain, Primary Neuron

SUBMITTER: Peter Nemes

LAB HEAD: Peter Nemes

PROVIDER: PXD028040 | Pride | 2022-02-18

REPOSITORIES: Pride

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