# High-throughput methods for measuring cortical synaptic connectivity at single-cell resolution

> **NIH NIH DP2** · BROAD INSTITUTE, INC. · 2022 · $900,000

## Abstract

Project Summary
 Information processing in the brain is accomplished by integrating neuronal activity via specific
patterns of synaptic connectivity between diverse neuronal subtypes, and changes in connectivity are
hypothesized to cause a range of neuropsychiatric disorders. However, current methods to measure
synaptic connectivity with single-cell precision are laborious, time-consuming, and costly. The main
goal of my research program is to develop high-throughput methods for measuring the patterns of
synaptic connectivity in the cerebral cortex. To accomplish this, I will pursue three separate approaches
using viral, functional, and molecular strategies. First, I will optimize rabies-based trans-synaptic tracing
to enable labeling of local cortical circuitry and classify connected neurons with multiplexed, error-
robust fluorescent in situ hybridization (merFISH). Next, I will screen for functional connectivity by
combining holographic optogenetic stimulation of individual pre-synaptic neurons with multiplexed
whole-cell automated patch-clamp to increase the number of synaptic connections that can be probed
from a single animal. Finally, I will design molecular barcodes targeted to pre- and post-synaptic sites
that can be imaged with merFISH to identify synaptic connections in concert with transcriptional
information for cell-type classification. I will first apply these methods towards high-confidence risk
genes associated with schizophrenia and bipolar disorder, as disordered cortical circuitry is thought to
cause these diseases. This proposal requires the innovative integration of multiple cutting-edge
technologies and the development of new, non-existing techniques. Given my training in
electrophysiology, optogenetics, advanced microscopy, and imaging analysis as well as my current
environment at the Stanley Center at the Broad Institute, I am ideally positioned to develop and validate
these techniques. Successful completion of this proposal will result in new methods for analyzing
synaptic connectivity applicable to a wide range of neuroscientific questions and provide insight into
the pathophysiological mechanisms of psychiatric disorders.

## Key facts

- **NIH application ID:** 10473009
- **Project number:** 1DP2MH132942-01
- **Recipient organization:** BROAD INSTITUTE, INC.
- **Principal Investigator:** Adam Granger
- **Activity code:** DP2 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $900,000
- **Award type:** 1
- **Project period:** 2022-09-01 → 2025-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10473009

## Citation

> US National Institutes of Health, RePORTER application 10473009, High-throughput methods for measuring cortical synaptic connectivity at single-cell resolution (1DP2MH132942-01). Retrieved via AI Analytics 2026-05-27 from https://api.ai-analytics.org/grant/nih/10473009. Licensed CC0.

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