Restoration of axon conduction and motor deficits by therapeutic treatment with glatiramer acetate.
Bottom Line: These GA-induced cytokines and growth factors may have a direct effect on axon function.Therapeutic regimens were utilized based on promising prophylactic efficacy.Finally, GA improved callosal axon conduction and nodal protein organization in EAE.
Affiliation: Department of Neurology, UCLA School of Medicine, Los Angeles, California.Show MeSH
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Mentions: Immunostaining was quantified using unbiased stereology. The dorsal column (DC) was delineated (Fig. 3A) using the drawing tool in ImageJ version 1.29 (Windows version of NIH Image; http://rsb.info/nih/gov/ij), and MBP, GFAP, CD3, and CD45 staining intensity was quantified within this region. NF200+ and MBP+ axons were counted in the ventral column (VC) of thoracic spinal cord (Fig. 3A). All images (RGB) were converted to gray scale, split, and separated by color channel in ImageJ. To avoid experimenter bias, autoadjustment of brightness, contrast, and threshold of staining signal were carried out in NIH software. A grid plug-in (ImageJ) was used for counting points per area of interest. Adenomatus polyposis coli (CC1)+ mature OLs, olig2+ OL lineage cells, and CD3+ T cells within the CC or spinal cord dorsal column were counted manually using ×10 or ×40 magnification images and compared blindly among normal, vehicle-treated EAE, and GA-treated EAE groups. Inflammatory cells were quantified by counting the number of CD45+ and CD3+ cells with DAPI+ nuclei in delineated thoracic spinal cord dorsal column (and/or delineated CC). Myelin (MBP+) and astrocytes (GFAP+) were calculated as percentage area intensity within the spinal cord dorsal column (Fig. 3) and delineated CC. Spinal cord axonal densities were calculated by counting the number of NF200+ cells in a ×40 image of ventral column of thoracic spinal cords, where coherent and similar diameter axons are present. Myelinated axon densities were calculated by counting axons (NF200+) with a clear ring of MBP+ myelin staining around them. Damaged axons were calculated by counting APP+ axons.
Affiliation: Department of Neurology, UCLA School of Medicine, Los Angeles, California.