Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2019 Dec 20;9(24):e3453.
doi: 10.21769/BioProtoc.3453.

Myelin Oligodendrocyte Glycoprotein 35-55 (MOG 35-55)-induced Experimental Autoimmune Encephalomyelitis: A Model of Chronic Multiple Sclerosis

Sakie Miyamura  1 , Nagisa Matsuo  1 , Kazuki Nagayasu  1 , Hisashi Shirakawa  1 , Shuji Kaneko  1
Affiliations

Myelin Oligodendrocyte Glycoprotein 35-55 (MOG 35-55)-induced Experimental Autoimmune Encephalomyelitis: A Model of Chronic Multiple Sclerosis

Sakie Miyamura et al. Bio Protoc. .

Abstract

Multiple sclerosis (MS) is the common demyelinating disease of human central nervous system. Among mouse models available to study MS, including the cuprizone application and lysolecithin-injection models, experimental autoimmune encephalomyelitis (EAE) model is widely used so that chronic EAE model of C57BL/6J can reflect the autoimmune pathogenesis of MS well. Here we introduce the EAE model based on C57BL/6J mice, which is generated by injection of myelin oligodendrocyte glycoprotein 35-55 (MOG 35-55) as an antigen. After immunization with complete Freund's adjuvant, clinical signs and changes in body weight are observed one or two weeks later. The EAE model will continue to be useful for development of therapeutics for MS.

Keywords: Autoimmune; C57BL/6J; Central nervous system (CNS); Chronic; Complete Freund's adjuvant (CFA); Experimental autoimmune encephalomyelitis (EAE); Multiple sclerosis (MS); Myelin oligodendrocyte glycoprotein 35-55 (MOG 35-55).

PubMed Disclaimer

Conflict of interest statement

Competing interestsThe authors declare no conflicts of interest associated with this manuscript.

Figures

Figure 1.
Figure 1.. Preparation of Complete Freund's Adjuvant (CFA).
A. Grounding MT with a mortar and a pestle. B. Mixing MT and 1 mg/ml CFA with a mortar and a pestle to obtain a final concentration of 6 mg/ml CFA/MT mix.
Figure 2.
Figure 2.. Preparation of the MOG emulsion.
A. Drawing 6 mg/ml CFA/MT mix from a mortar into a 2.5 ml leur lock syringe. B. Connecting CFA syringe to a three-way stopcock and releasing air from the stopcock. C. Closing the valve connected to the CFA syringe. D. Connecting MOG 35-55 syringe to the stopcock and releasing air. E. Arranging amount of MOG 35-55:CFA = 1:1 and closing the valve without syringe. F. Mixing MOG 35-55 and CFA to get white emulsion.
Figure 3.
Figure 3.. Checking the condition of the emulsion.
A. Unfinished emulsion spread like this picture. B. Completed emulsion does not spread.
Figure 4.
Figure 4.. Animal immunization.
A. Connecting a 26 G needle to the emulsion syringe. B and C. Injecting emulsion subcutaneously to the neck (B) and base of the right hindlimb (C).
Figure 5.
Figure 5.. Clinical score evaluation of EAE mice.
Representative mouse images of each clinical score. A. Clinical score 1: partial tail paralysis, B. Clinical score 2: full tail paralysis, C. Clinical score 3: partial hindlimb paralysis, D. Clinical score 4: full hindlimb paralysis, E. Clinical score 5: forelimb paresis.
Video 1.
Video 1.. EAE mouse with clinical score 1
(All experiments were conducted in accordance with the ethical guidelines of the Kyoto University Animal Experimentation Committee and with the guidelines of the Japanese Pharmacological Society. The approval ID of the animal experiment in this protocol is 14-42 and the validity period is from 2014 to 2019.(
Video 2.
Video 2.. EAE mouse with clinical score 2
(All experiments were conducted in accordance with the ethical guidelines of the Kyoto University Animal Experimentation Committee and with the guidelines of the Japanese Pharmacological Society. The approval ID of the animal experiment in this protocol is 14-42 and the validity period is from 2014 to 2019.(
Video 3.
Video 3.. EAE mouse with clinical score 3
(All experiments were conducted in accordance with the ethical guidelines of the Kyoto University Animal Experimentation Committee and with the guidelines of the Japanese Pharmacological Society. The approval ID of the animal experiment in this protocol is 14-42 and the validity period is from 2014 to 2019.(
Video 4.
Video 4.. EAE mouse with clinical score 4
(All experiments were conducted in accordance with the ethical guidelines of the Kyoto University Animal Experimentation Committee and with the guidelines of the Japanese Pharmacological Society. The approval ID of the animal experiment in this protocol is 14-42 and the validity period is from 2014 to 2019.(
Video 5.
Video 5.. EAE mouse with clinical score 5
(All experiments were conducted in accordance with the ethical guidelines of the Kyoto University Animal Experimentation Committee and with the guidelines of the Japanese Pharmacological Society. The approval ID of the animal experiment in this protocol is 14-42 and the validity period is from 2014 to 2019.(
Figure 6.
Figure 6.. Changes in weight and clinical symptoms of EAE.
Development of EAE in MOG-immunized C57BL/6 mice. Body weight (A) and clinical score (B) were monitored for 28 days. Note that mice began to lose body weight at the onset of clinical signs during EAE. n = 4, **P < 0.01, ***P < 0.001.
See this image and copyright information in PMC

References

    1. Baker D. and Amor S.(2014). Experimental autoimmune encephalomyelitis is a good model of multiple sclerosis if used wisely. Mult Scler Relat Disord 3(5): 555-564. - PubMed
    1. Bittner S., Afzali A. M., Wiendl H. and Meuth S. G.(2014). Myelin oligodendrocyte glycoprotein(MOG35-55) induced experimental autoimmune encephalomyelitis(EAE) in C57BL/6 mice. J Vis Exp (86). - PMC - PubMed
    1. Burrows D. J., McGown A., Jain S. A., De Felice M., Ramesh T. M., Sharrack B. and Majid A.(2019). Animal models of multiple sclerosis: From rodents to zebrafish. Mult Scler 25(3): 306-324. - PubMed
    1. Gold R., Linington C. and Lassmann H.(2006). Understanding pathogenesis and therapy of multiple sclerosis via animal models: 70 years of merits and culprits in experimental autoimmune encephalomyelitis research. Brain 8): 1953-1971. - PubMed
    1. Hall S. M.(1972). The effect of injections of lysophosphatidyl choline into white matter of the adult mouse spinal cord. J Cell Sci 10(2): 535-546. - PubMed

LinkOut - more resources