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HMGB1 Contributes to Regeneration After Spinal Cord Injury in Adult Zebrafish

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Abstract

High mobility group box 1 (HMGB1, also called amphoterin) facilitates neurite outgrowth in early development, yet can exacerbate pathology and inhibit regeneration by inducing adverse neuroinflammation when released from dying cells, suggesting that HMGB1 plays a critical, yet undefined role in neuroregeneration. We explored whether HMGB1 contributes to recovery after complete spinal cord transection in adult zebrafish. Quantitative PCR and in situ hybridization revealed that HMGB1 mRNA levels decreased between 12 h to 11 days after spinal cord injury (SCI), then returned to basal levels by 21 days. Western blot and immunohistological analyses indicated that the time course of HMGB1 protein expression after SCI parallels that of mRNA. Immunofluorescence staining revealed that HMGB1 translocates from nuclei into the cytoplasm of spinal motoneurons at 4 and 12 h (acute stage) following SCI, then accumulates in the nuclei of motoneurons during the ensuing chronic stage (after 6 days following SCI). Immunohistology of transgenic zebrafish, expressing green fluorescent protein in blood vessels, showed enhanced HMGB1 expression in blood vessels in the vicinity of motoneurons. Application of anti-sense HMGB1 morpholinos inhibited locomotor recovery by 34 % and decreased axonal regeneration by 34 % compared to fish treated with a control morpholino. The present study shows that HMGB1 expression increases in both endothelial cells and motoneurons, suggesting that HMGB1 promotes recovery from SCI not only through enhancing neuroregeneration, but also by increasing angiogenesis. The inflammatory effects of HMGB1 are minimized through the decrease in HMGB1 expression during the acute stage.

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Acknowledgments

This project is supported by the National Natural Science Foundation of China (No. 81072622, No. 31271580), Guangdong Natural Science Foundation (S2012010009480) and the Li Kashing Foundation. Melitta Schachner is New Jersey Professor of Spinal Cord Research. We are grateful to Zilong Wen for kindly providing the Fli1a::EGFP transgenic fish.

Conflicts of Interest

The authors declare that there are no conflicts of interest.

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Author notes

  1. Yan-Qin Shen

    Present address: Medical School, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, China

Authors and Affiliations

  1. Center for Neuroscience, Shantou University Medical College, 22 Xinling Road, Shantou, 515041, China

    Ping Fang, Hong-Chao Pan, Stanley Li Lin, Melitta Schachner & Yan-Qin Shen

  2. Key Laboratory of Zebrafish Modeling and Drug Screening for Human Diseases of Guangdong Higher Education Institutes, Department of Cell Biology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, 510515, China

    Wen-Qing Zhang

  3. Neuroscience Center, University of Helsinki, P.O. Box 56 (Viikinkaari 4), FIN-00014, Helsinki, Finland

    Heikki Rauvala

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  1. Ping Fang
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Correspondence to Melitta Schachner or Yan-Qin Shen.

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Fang, P., Pan, HC., Lin, S.L. et al. HMGB1 Contributes to Regeneration After Spinal Cord Injury in Adult Zebrafish. Mol Neurobiol 49, 472–483 (2014). https://doi.org/10.1007/s12035-013-8533-4

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  • DOI: https://doi.org/10.1007/s12035-013-8533-4

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