A method for histopathological study of the multifocal nature of spinal cord lesions in murine experimental autoimmune encephalomyelitis

PeerJ

article

Katherine N. Gibson-Corley¹  Alexander W. Boyden¹  Mariah R. Leidinger¹  Allyn M. Lambertz¹  Georgina Ofori-Amanfo¹  Paul W. Naumann¹  J. Adam Goeken¹  Nitin J. Karandikar¹ 

[1] Department of Pathology, University of Iowa

关键词: Mice;    Histopathology;    Spinal cord;    EAE;   

DOI  :  10.7717/peerj.1600

学科分类：社会科学、人文和艺术（综合）

来源: Inra

【 摘 要 】

Experimental autoimmune encephalomyelitis (EAE) is a well-established mouse model for multiple sclerosis and is characterized by infiltration of mononuclear cells and demyelination within the central nervous system along with the clinical symptoms of paralysis. EAE is a multifocal and random disease, which sometimes makes histopathologic analysis of lesions difficult as it may not be possible to predict where lesions will occur, especially when evaluating cross sections of spinal cord. Consequently, lesions may be easily missed due to limited sampling in traditional approaches. To evaluate the entire length of the spinal cord while maintaining anatomic integrity, we have developed a method to section the cord within the decalcified spinal column, which allows for the study of the multifocal nature of this disease and also minimizes handling artifact. HE and Luxol fast blue staining of these spinal cord sections revealed a paucity of lesions in some areas, while others showed marked inflammation and demyelination. The percentage of spinal cord affected by EAE was evaluated at four separate areas of longitudinally sectioned cord and it varied greatly within each animal. Immunohistochemical staining of in situ spinal cords which had undergone decalcification was successful for key immuno-markers used in EAE research including CD3 for T cells, B220 for B cells and F4/80 for murine macrophages. This method will allow investigators to look at the entire spinal cord on a single slide and evaluate the spinal cord with and without classic EAE lesions.

【 授权许可】

CC BY