Abstract
After skeletal muscle injury, unloading disturbs the regenerative process of injured myofibers, in a manner highly attributed to impairment of macrophage functions. However, the effect of unloading on the spatiotemporal context of proinflammatory macrophage recruitment and satellite cell accumulation within the damaged area remains unclear. This study focused on macrophages expressing inducible nitric oxide synthase (iNOS) that synthesize nitric oxide, a key regulator of muscle regeneration, and compared the continuous hindlimb unloading (HU) by tail suspension versus weight-bearing (WB) after skeletal muscle crush injury in rats. We found that in the WB group, the recruitment of iNOS+ proinflammatory macrophages into the injured site gradually increased until their peak number at 48 h post-injury. In the HU group, the accumulation of iNOS+ macrophages until 48 h after injury was significantly less than that in the WB group and continued to increase at 72 h. In accordance with attenuated and/or delayed iNOS+ macrophage recruitment, whole iNOS expression at 24 and 48 h after injury was weakened by unloading. Additionally, in the HU group, satellite cell content of dystrophin-positive non-injured areas diminished at 48 h after injury, and the numbers of activated satellite cells within the regenerating area at 72 and 96 h post-injury were significantly smaller than those in the WB group. These findings suggest that muscle regeneration under unloading conditions results in attenuated and/or delayed recruitment of iNOS+ macrophages and lower iNOS expression in the early phase after muscle injury, leading to perturbed satellite cell accumulation and muscle regeneration.
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Acknowledgments
Authors thank the members of our laboratory for their cooperation. We also thank ENAGO (https://www.enago.jp/) for English language editing. This work was supported by Japan Society for the Promotion of Science Grant-in-Aid for scientific research KAKENHI No. 17K01501.
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Conceptualization: MK and TA; Methodology: MK, MM, MS and TA; Formal analysis and investigation: MK, MM, MS and MM; Writing—original draft preparation: MK; Writing—review and editing: MM, MS and TA; Funding acquisition: TA. All authors read and approved the final manuscript.
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Kawashima, M., Miyakawa, M., Sugiyama, M. et al. Unloading during skeletal muscle regeneration retards iNOS-expressing macrophage recruitment and perturbs satellite cell accumulation. Histochem Cell Biol 154, 355–367 (2020). https://doi.org/10.1007/s00418-020-01897-3
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DOI: https://doi.org/10.1007/s00418-020-01897-3