A Simple Approach to Induce Experimental Autoimmune Neuritis in C57BL/6 Mice for Functional and Neuropathological Assessments

David G. Gonsalvez; Jessica L. Fletcher; Sang Won Yoo; Rhiannon J. Wood; Simon S. Murray; Junhua Xiao

doi:10.3791/56455

A Simple Approach to Induce Experimental Autoimmune Neuritis in C57BL/6 Mice for Functional and N...

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JoVE Journal Neuroscience

A Simple Approach to Induce Experimental Autoimmune Neuritis in C57BL/6 Mice for Functional and Neuropathological Assessments

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07:30 min

November 9, 2017

DOI: 10.3791/56455-v

David G. Gonsalvez¹, Jessica L. Fletcher¹, Sang Won Yoo¹, Rhiannon J. Wood¹, Simon S. Murray¹, Junhua Xiao¹

¹Department of Anatomy and Neuroscience, School of Biomedical Sciences, Faculty of Medicine, Dentistry and Health Sciences,The University of Melbourne

Overview

This study presents a methodology to induce Experimental Autoimmune Neuritis (EAN) in a mouse model using the myelin protein zero (P0) 180-199 peptide along with Freund's complete adjuvant and pertussis toxin. The aim is to facilitate a functional and pathological analysis of peripheral neuropathy, providing insights into the disease process and potential treatments.

Key Study Components

Area of Science

Neuroscience
Autoimmune disorders
Peripheral neuropathy

Background

Experimental Autoimmune Neuritis (EAN) is a model for studying peripheral neuropathy.
The model is practical for assessing new therapeutic agents.
Functional testing during the study aids in objective evaluation.
Key behavior assessments include motor function and clinical scoring.

Purpose of Study

To induce EAN efficiently in C57BL/6 mice for research purposes.
To analyze the extent of functional deficits and neuropathology.
To establish a reproducible protocol for future studies on peripheral neuropathy.

Methods Used

The platform involves using a treadmill and gait function imaging system for functional assessments.
The model focuses on male C57BL/6 mice for the EAN induction process.
No multiomics workflows were mentioned in the study.
The protocol includes various subcutaneous injections at specified intervals post-immunization.
Motor function assessments are carried out multiple times during the study to monitor disease progression.

Main Results

The study confirms a monophasic disease course with clinical symptoms peaking 25 days after immunization.
Significant functional declines in motor task performance are observed from day six to day 35.
Neuropathological analysis shows demyelination and axonal damage in treated mice compared to controls.
Elevated beta-amyloid precursor protein expression indicates acute axonal injury.

Conclusions

This study provides a clear and efficient protocol for inducing EAN, facilitating understanding of neuropathology.
Insights into the disease process support future research on potential treatments for peripheral neuropathy.
These findings enhance comprehension of autoimmune mechanisms affecting the peripheral nervous system.

Frequently Asked Questions

What are the advantages of this EAN model?

The EAN model allows for the objective assessment of functional and pathological changes associated with peripheral neuropathy, making it valuable for therapeutic investigations.

How is the EAN induction implemented?

EAN is induced through subcutaneous injections of a mixture of peptide and adjuvant, along with pertussis toxin, in C57BL/6 mice.

What types of data are obtained from this study?

Key outcomes include motor function assessments, clinical scores, and neuropathological evaluations, which provide insights into disease progression.

Can this method be adapted for different models?

While primarily designed for C57BL/6 mice, the protocol may be adapted for other strains with consideration for their specific immune responses.

What are some limitations of this approach?

The primary limitation is the specificity of the model to induce mono-phasic EAN, which may not fully represent other forms of neuropathy.

This report outlines a simple approach to successfully induce experimental autoimmune neuritis (EAN) using the myelin protein zero (P0)_180-199 peptide in combination with Freund's complete adjuvant and pertussis toxin. We present a sophisticated paradigm capable of accurately assessing the extent of functional deficits and neuropathology that occur in this EAN.

The overall goal of this procedure is to successfully induce Experimental Autoimmune Neuritis or EAN in a mouse model for the functional and pathological analysis of the disease process. The EAN model can be used to address key questions relating to peripheral neuropathy. It's also a wonderful model to potentially investigate the efficacy of novel therapeutic agents.

The main advantage of this approach is that it provides a simple and objective functional testing paradigm that can be applied when using the EAN model in the mouse. Demonstrating the procedure with me today is Sang Won Yoo, who is an undergraduate student in the neurotrophin and myelin biology lab at the University of Melbourne. Three days before the disease induction, turn on the motor function assessment apparatus and switch on the light button.

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