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. 2020 Aug 4;10:13138. doi: 10.1038/s41598-020-70068-0

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Motor deficits and histopathologies of a rat model of CSM. (A) A representative T2-weighted mid-sagittal MRI image of a CSM rat at 10 weeks after surgery shows dorsal–ventral cervical spinal cord compression at C5 and C6 levels while keeping laminae of C4, 5, 6, and 7 intact. The area in the red dotted square is schematically shown in the inset. (B) Upper panels schematically show the spinal cord around the C5-6 level. Hematoxylin and eosin (HE) staining of axial sections at the red broken lines at the C5-6 disc level are shown in the lower panels. Blue boxed areas are enlarged in (E). Bar = 600 μm. (C,D) Stride length (C) and duty cycle (D) of fore- and hind-limbs analyzed by the CatWalk system of sham-operated and CSM rats from 1 to 10 weeks postoperatively. (C) A representative stride length of the right forelimb is indicated by a double-headed arrow in the right panel. (D) Duty cycle is a ratio in time when a paw stays on the glass plate in a single step cycle. CSM rats started to exhibit gait disturbance from 5 weeks after surgery. Mean and SE are indicated (n = 6 and 4 for sham-operated and CSM rats, respectively). *p < 0.05, **p < 0.01, and ***p < 0.001 by two-way repeated measures ANOVA followed by Bonferroni correction. (E,F) HE staining at 5 weeks postoperatively (E) and 4-hydroxynonenal (4-HNE) staining at 10 weeks postoperatively (F) of an axial section of the spinal cord at the C5-6 disc level in sham-operated and CSM rats. (E) Images are enlargement of the blue boxed areas in (B). A representative neuron (white arrow) in the anterior horn (AH) region is enlarged in the inset. Purple-stained Nissl bodies are indicated by yellow arrowheads. The numbers of cells with Nissl bodies in the AH region of sham-operated and CSM rats are shown. Mean and SD are indicated (n = 3 rats in each group). *p < 0.05 by Mann–Whitney U-test. (F) Representative images of the spinal cord stained with 4-hydroxynonenal (4-HNE). Arrows point to 4-HNE-positive large cells (diameter > 40 μm), which were recognized as motor neurons, in the AH region. A border between the gray and the white matters is indicated by a red dotted line. Bar = 50 μm. Mean and SD are indicated (n = 3 rats in each group). *p < 0.05 by Mann–Whitney U-test.