Neural Regeneration Research ›› 2021, Vol. 16 ›› Issue (12): 2549-2556.doi: 10.4103/1673-5374.313052

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Spastin interacts with collapsin response mediator protein 3 to regulate neurite growth and branching

Zhi-Sheng Ji1, #, Jian-Ping Li2, #, Chao-Hua Fu1, 3, #, Jian-Xian Luo1, Hua Yang1, Guo-Wei Zhang1, Wutian Wu2, 4, 5, *, Hong-Sheng Lin1, *   

  1. 1Department of Orthopedics, The First Affiliated Hospital of Jinan University, Guangzhou, Guangdong Province, China; 2Guangdong-Hong Kong-Macau Institute of CNS Regeneration, Jinan University, Guangzhou, Guangdong Province, China; 3Department of Orthopedics, Jiangmen Hospital of Sun Yat-sen University, Jiangmen, Guangdong Province, China; 4Re-Stem Biotechnology Co., Ltd., Suzhou, Jiangsu Province, China; 5Spine Surgery, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong Province, China
  • Online:2021-12-15 Published:2021-05-17
  • Contact: Wutian Wu, MD, PhD, wtwu@hkucc.hku.hk; Hong-Sheng Lin, MD, PhD, tlinhsh@jnu.edu.cn.
  • Supported by:
    This work was supported by the National Natural Science Foundation of China, Nos. 31900691 (to GWZ), 81771331 (to HSL) and 81971165 (to WW); the National Basic Research Program of China (973 Program), No. 2014CB542205 (to WW); the Natural Science Foundation of Guangdong Province of China, No. 2017A030313595 (to HSL); the Science and Technology Program of Guangzhou, China, No. 201707010370 (to HSL); Project of Educational Commission of Guangdong Province of China, No. 2018KQNCX013 (to ZSJ); the Fundamental Research Funds for the Central Universities Project, China, No. 21618304 (to GWZ); Guangdong Provincial Key Research and Development Program “Precision Medicine and Stem Cell” Major Science and Technology Project, China, No. 3242001 (to WW); and China Postdoctoral Science Foundation, No. 2019M653292 (to ZSJ).

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Abstract: Cytoskeletal microtubule rearrangement and movement are crucial in the repair of spinal cord injury. Spastin plays an important role in the regulation of microtubule severing. Both spastin and collapsin response mediator proteins can regulate neurite growth and branching; however, whether spastin interacts with collapsin response mediator protein 3 (CRMP3) during this process remains unclear, as is the mechanism by which CRMP3 participates in the repair of spinal cord injury. In this study, we used a proteomics approach to identify key proteins associated with spinal cord injury repair. We then employed liquid chromatography-mass spectrometry to identify proteins that were able to interact with glutathione S-transferase-spastin. Then, co-immunoprecipitation and staining approaches were used to evaluate potential interactions between spastin and CRMP3. Finally, we co-transfected primary hippocampal neurons with CRMP3 and spastin to evaluate their role in neurite outgrowth. Mass spectrometry identified the role of CRMP3 in the spinal cord injury repair process. Liquid chromatography-mass spectrometry pulldown assays identified three CRMP3 peptides that were able to interact with spastin. CRMP3 and spastin were co-expressed in the spinal cord and were able to interact with one another in vitro and in vivo. Lastly, CRMP3 overexpression was able to enhance the ability of spastin to promote neurite growth and branching. Therefore, our results confirm that spastin and CRMP3 play roles in spinal cord injury repair by regulating neurite growth and branching. These proteins may therefore be novel targets for spinal cord injury repair. The Institutional Animal Care and Use Committee of Jinan University, China approved this study (approval No. IACUS-20181008-03) on October 8, 2018. 

Key words: collapsin response mediator protein 3, liquid chromatography-mass spectrometry, microtubule, neurite growth, protein interactions, proteomics, spastin, spinal cord injury

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