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. 2021 May 21;18:117. doi: 10.1186/s12974-021-02168-1

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© The Author(s) 2021

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 1 — CCI evokes increases in the expressions of CGRP and Iba1 in the L4L5 spinal dorsal horn of CCI rats. a Nociceptive behavior developed in CCI model rats. Mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) were examined at 0, 1, 3, 5, 7, 10, and 14 days respectively after sham operation or CCI surgery. n = 6, ^*p < 0.05 vs. sham group. b Double-staining immunofluorescent images showing the CGRP-positive fibers (green) and Iba1 (microglia maker)-positive microglia (red) in the dorsal horn of sham and CCI groups on day 5 after surgery. Cell nuclei were stained with the DAPI (blue). Note that numerous varicose nerve terminals immunoreactive for CGRP (green) closely approached and surrounded Iba1 immunopositive microglia (red) in the laminae I and II of the spinal dorsal horn (second row). Scale bar of 200 m in the first row and 10 m in the second row. c Quantitative analysis of the percentages of CGRP-immunoreactive surface in laminae I and II, and Iba1-immunostaining surface in the spinal dorsal horn showed the CCI-induced changes. Data are presented as the mean SEM (n = 6). ^*p < 0.05 vs. sham group. d Western blot analysis of CGRP expression in the spinal dorsal horn on 0, 1, 3, 5, 7, 10, and 14 days after CCI surgery, respectively. The mean optic density of the protein was calculated by normalizing to GAPDH. All values are expressed as the means SEMs (n = 4).^*p < 0.05 vs. sham group