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. 2017 Aug 21;7:9018. doi: 10.1038/s41598-017-09432-6

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© The Author(s) 2017

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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5-HT+ nerve fibers fail to regenerate in the chronically repaired cord. (a–d) Images are from PNG+aFGF+ChABC-treated animals. The dotted line in each panel delineates the interface between the graft and spinal cord. (a) A representative low-magnification confocal image of a sagittal section with GFAP labeling (green) and 5-HT staining (red) displaying the overall anatomy of the PNG, spinal cord, and regenerating 5-HT+ axons. Scale bar, 500 µm. (b) High-magnification confocal image of the rostral cord/PNG interface boxed in a. Note the small density of 5-HT+ stained axons (red) entering the PNG (GFAP-negative area) from the rostral end of the spinal cord (green). Scale bar, 100 µm. (c) High-magnification confocal image of the caudal PNG/cord interface boxed in a. Few 5-HT+ stained axons (red) reach the distal end of the graft, and no 5-HT axons exit the graft back into the caudal spinal cord (green). Scale bar, 100 µm. (d) Low-magnification confocal image of a sagittal section with GFAP labeling (green) and SERT staining (red) displaying the overall anatomy of the PNG, spinal cord, and lack of regenerating SERT+ axons. Scale bar, 500 µm. (e–g) Images are from PNG-treated animals. The dotted line in each panel delineates the interface between the graft and spinal cord. (e) A representative low-magnification confocal image of a sagittal section with GFAP labeling (green) and 5-HT staining (red) displaying the overall anatomy of the PNG, spinal cord, and regenerating 5-HT+ axons. Scale bar, 500 µm. (f) High-magnification confocal image of the rostral cord/PNG interface boxed in e. Note sparse 5-HT+ stained axons (red) entering the PNG (GFAP-negative area) from the rostral end of the spinal cord (green). Scale bar, 100 µm. (g) High-magnification confocal image of the caudal PNG/cord interface boxed in e. Note that no 5-HT+ stained axons (red) reach the distal end of the graft and no 5-HT+ axons exit the graft back into the caudal spinal cord (green). Scale bar, 100 µm. (h) Quantification of the density of 5-HT-immunoreactive fibers found in the rostral spinal cord adjacent to the PNG, in the center of the PNG, and in the caudal spinal cord adjacent to the distal PNG. n = 6 per group. *p < 0.05, Student’s t-test. Data represent mean ± S.E.M.