# Determining structure and organization of neurofilaments in situ using cryo-electron tomography

> **NIH NIH R21** · BAYLOR COLLEGE OF MEDICINE · 2021 · $46,805

## Abstract

Project Summary
As one of the major components of the neuronal cytoskeleton network, neurofilaments are essential
for the radial growth and conduction velocity of axons. Neurofilaments are involved in the
pathogenesis of multiple neurological disorders and serve as important axonal damage biomarkers in
neurodegenerative and traumatic diseases. Despite their critical role in the structure and function of
axons, little is known about the molecular assembly of neurofilaments. Currently, it is still unclear how
individual neurofilament subunits assemble into mature filaments, or how those filaments interact with
each other to form the cytoskeleton network that supports the growth of axons.
Cryo-electron tomography (CryoET) allows visualization of protein complexes inside cells at
nanometer resolution in three dimensions. Detailed structural information of those proteins can be
obtained from the tomogram using specialized computational techniques. In this proposal, we aim to
study the structure and organization of neurofilaments within axons using CryoET.
To achieve this goal, we will image the axon of primary neurons with CryoET and determine the
structures of neurofilaments at subnanometer resolution using subtomogram classification and
averaging. Molecular models will be constructed from the averaged structures to reveal the in situ
assembly of neurofilaments. The structures and models of neurofilaments will be studied in their
cellular context to understand how the neuronal cytoskeleton organization evolves, as the axons grow
thicker. To make the proposed work possible, novel computational methods will be developed for
CryoET data analysis throughout the process.
We believe this research will provide the first structural insight into in situ neurofilament assembly and
deepen our understanding of neuronal cytoskeleton organization. The structural information acquired
from this research will pave the way for future studies of the axon maturation process, as well as the
structural basis of various neurological diseases.

## Key facts

- **NIH application ID:** 10303416
- **Project number:** 1R21MH125285-01A1
- **Recipient organization:** BAYLOR COLLEGE OF MEDICINE
- **Principal Investigator:** Muyuan Chen
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $46,805
- **Award type:** 1
- **Project period:** 2021-07-01 → 2022-01-09

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10303416

## Citation

> US National Institutes of Health, RePORTER application 10303416, Determining structure and organization of neurofilaments in situ using cryo-electron tomography (1R21MH125285-01A1). Retrieved via AI Analytics 2026-05-26 from https://api.ai-analytics.org/grant/nih/10303416. Licensed CC0.

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