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.