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Updated: May 28, 2026
Coherent Anti-Stokes Raman Spectroscopy (CARS) Application for Imaging Myelination in Brain Slices
Published on: July 22, 2022
Paranodal myelin retraction in relapsing experimental autoimmune encephalomyelitis visualized by coherent anti-Stokes
Yan Fu1, Terra J Frederick, Terry B Huff
1Purdue University, Weldon School of Biomedical Engineering, 206 South Martin Jischke Drive, West Lafayette, Indiana 47907, USA.
Abstract:
How demyelination is initiated is a standing question for pathology of multiple sclerosis. By label-free coherent anti-Stokes Raman scattering (CARS) imaging of myelin lipids, we investigate myelin integrity in the lumbar spinal cord tissue isolated from naïve SJL mice, and from mice at the onset, peak acute, and remission stages of relapsing experimental autoimmune encephalomyelitis (EAE). Progressive demyelinating disease is initially characterized by the retraction of paranodal myelin both at the onset of disease and at the borders of acute demyelinating lesions. Myelin retraction is confirmed by elongated distribution of neurofascin proteins visualized by immunofluorescence. The disruption of paranodal myelin subsequently exposes Kv1.2 channels at the juxtaparanodes and lead to the displacement of Kv1.2 channels to the paranodal and nodal domains. Paranodal myelin is partially restored during disease remission, indicating spontaneous myelin regeneration. These findings suggest that paranodal domain injury precedes formation of internodal demyelinating lesions in relapsing EAE. Our results also demonstrate that CARS microscopy is an effective readout of myelin disease burden.
Insights
Investigating multiple sclerosis pathology, this study reveals that paranodal myelin injury precedes demyelination in relapsing experimental autoimmune encephalomyelitis (EAE). Myelin regeneration occurs during remission, suggesting CARS microscopy as a valuable tool for assessing myelin disease burden.
Area of Science:
- Neuroscience
- Immunology
- Pathology
Background:
- The initiation of demyelination in multiple sclerosis (MS) remains a critical question in its pathology.
- Understanding myelin integrity is crucial for developing effective MS treatments.
Purpose of the Study:
- To investigate myelin integrity during the course of relapsing experimental autoimmune encephalomyelitis (EAE) in mice.
- To determine the sequence of events in demyelination and identify potential therapeutic targets.
Main Methods:
- Label-free coherent anti-Stokes Raman scattering (CARS) microscopy was employed to image myelin lipids in mouse spinal cord tissue.
- Immunofluorescence staining was used to visualize neurofascin protein distribution.
Main Results:
- Paranodal myelin retraction was observed at the onset and borders of acute lesions in EAE mice.
- Disruption of paranodal myelin led to the displacement of Kv1.2 channels.
- Partial restoration of paranodal myelin during remission indicated spontaneous myelin regeneration.
- CARS microscopy proved effective in quantifying myelin disease burden.
Conclusions:
- Paranodal myelin injury precedes internodal demyelination in relapsing EAE.
- Spontaneous myelin regeneration occurs during disease remission.
- CARS microscopy is a valuable tool for assessing myelin pathology in MS models.
