# High Throughput Methods for Single Cell Proteomics

> **NIH NIH R21** · SCRIPPS RESEARCH INSTITUTE, THE · 2022 · $266,250

## Abstract

PROJECT SUMMARY/ ABSTRACT
To understand deviations in normal function of neurons and neuronal circuits brought on by disease, we
will develop methods using patch clamping to measure the electrophysiology of individual neurons and
then use mass spectrometry-based proteomics to measure the proteome. Now that neurons can be
created from the skin cells of patients harboring disease genes to determine genotype to phenotype
relationships between the genetic defects and electrophysiology, the methods we develop have great
potential to significantly improve our ability to study human diseases of the brain. “Brain” organoids are
also being created from skin cells to recapitulate a 3-dimensional environment for neurons and to include
excitatory and inhibitory neurons. The ability to concurrently measure electrophysiology and protein
expression will allow a determination of how disease related perturbations to neurons and other cells are
related to molecular phenotypes. Single cell RNA-SEQ is used to measure gene expression in neurons,
but gene expression profiles fail to account for rates of protein synthesis, degradation, proteostasis, post
translational modification and enzymatic activity, all of which are critical cellular functions accomplished
by proteins. Single cell mass spectrometry has been applied to neurons to measure metabolites and
neuropeptides, but efforts to measure the proteome have lagged. We have established that proteins can
be measured in neurons after electrophysiology, but here we propose to greatly increase the scale of
measurements as well as the throughput. These methods will be broadly applicable as patch clamping
techniques is a widely used technique to measure ionic currents in a variety of cell types including
neurons, cardiomyocytes, muscle fibers and pancreatic beta cells. Furthermore, these methods will
enable experiments to determine the mechanism of action of drugs that restore normal electrophysiology
to neurons

## Key facts

- **NIH application ID:** 10433158
- **Project number:** 1R21MH129776-01
- **Recipient organization:** SCRIPPS RESEARCH INSTITUTE, THE
- **Principal Investigator:** John R Yates III
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $266,250
- **Award type:** 1
- **Project period:** 2022-05-01 → 2024-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10433158

## Citation

> US National Institutes of Health, RePORTER application 10433158, High Throughput Methods for Single Cell Proteomics (1R21MH129776-01). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10433158. Licensed CC0.

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