# Electrified cryo-EM: a new tool to capture metastable neuron structures during an action potential

> **NIH NIH DP2** · UNIVERSITY OF CALIFORNIA LOS ANGELES · 2024 · $1,417,500

## Abstract

Project Summary:
There remains a critical blind spot in the study of biological systems. In situ imaging techniques like super-resolution
microscopy can elucidate cellular dynamics but cannot provide the sub-nanometer resolution needed for molecular
detail. Cryogenic electron microscopy (cryo-EM) does provide atomic-resolution structures yet remains limited to
only systems in their equilibrium states. While both these techniques have been awarded separate Nobel prizes for
their transformative impact on our understanding of biology, a missing gap still exists for how the structure of
biomolecules change in their active state far from equilibrium. The proposed research seeks to bridge this gap. This
project will pioneer the development of electrified cryo-EM (eCryo-EM), a novel tool developed by my research
group that can kinetically trap biological systems in their metastable state away from equilibrium. Briefly, an electrical
stimulus applied just prior to and throughout a plunge-freezing step will capture and preserve metastable states in
electrically excitable biological systems. This approach in using eCryo-EM to capture metastable state is general for
all of biology. Indeed, many fundamental questions remain unanswered for a broad spectrum of electrically excitable
biological systems with important implications for medical applications (e.g., wound healing, electroporation drug
delivery, muscle contraction, sensory acquisition, etc.). As a starting point, this proposal will unravel the first snapshots
of neuron conformational changes through the stages of an action potential across multiple length scales. At the
molecular level, eCryo-EM will reveal the molecular structure of voltage-gated channel proteins as they change
through open, closed, and inactivated states. At the cell level, eCryo-EM will reveal the mechanism of neural inhibition
at the synaptic cleft under high-frequency waveforms. These new insights not only provide fundamental
understanding for neuron behavior under an electrical stimulus, but also will guide future neurostimulation treatments
that are directed and tailored for a specific neural circuit to restore human health in patients suffering from
neurological disease. 3 tasks are outlined to achieve the stated goals: (1) demonstration of an eCryo-EM device that
can successfully trap metastable states of biological systems under an electrical stimulus; (2) elucidating a time series
of structural changes in voltage-gated channel proteins throughout an action potential; and (3) trapping the multiple
conformations of a synaptic junction between neurons throughout electrical stimulation. These tasks cannot be
achieved through conventional methodologies. Rather, it requires a fundamentally new approach in kinetically
trapping the active state biological systems directly under an electrical bias. The proposed research will establish a new
paradigm in investigating how neurons fundamentally behave in response to an external electr...

## Key facts

- **NIH application ID:** 10910505
- **Project number:** 1DP2NS142715-01
- **Recipient organization:** UNIVERSITY OF CALIFORNIA LOS ANGELES
- **Principal Investigator:** Yuzhang Li
- **Activity code:** DP2 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $1,417,500
- **Award type:** 1
- **Project period:** 2024-09-06 → 2027-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10910505

## Citation

> US National Institutes of Health, RePORTER application 10910505, Electrified cryo-EM: a new tool to capture metastable neuron structures during an action potential (1DP2NS142715-01). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10910505. Licensed CC0.

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