title: Involvement of gecko SNAP25b in spinal cord regeneration by promoting outgrowth and elongation of neurites.
abstract: SNARE complex mediates cellular membrane fusion events essential for neurotransmitter release and synaptogenesis. SNAP25, a member of the SNARE proteins, plays critical roles during the development of the central nervous system via regulation by alternative splicing and protein kinase phosphorylation. To date, little information is available regarding the protein in the spinal cord regeneration, especially for the postnatal highly expressed isoform SNAP25b. In the present study, we characterized gecko SNAP25b, which shared high identity with those of other vertebrates. Expression of gecko SNAP25b was temporally upregulated in both neurons of spinal cord and forming ependymal tube following tail amputation, coinciding with the occurrence of regenerate re-innervation. Overexpression of gecko wild type SNAP25b in the SH-SY5Y and undifferentiated PC12 cells promoted the elongation and outgrowth of neurites, while mutant constructs at Serine(187) resulted in differential effects for which S187A had a promoting role. Knockdown of endogenous SNAP25b affected the formation of neurites, which could be rescued by overexpression of SNAP25b. FM1-43 staining revealed that transfection of S187E mutant construct reduced the recruitment of vesicles. In addition, transfection of gecko SNAP25b in the astrocyte, which is absent from neuronal specific VAMP2, was capable of enhancing process elongation, indicating a potential for various alternative protein combinations. Taken together, our data suggest that gecko SNAP25b is involved in spinal cord regeneration by promoting outgrowth and elongation of neurites in a more extensive protein binding manner.
authors.affiliation: Key Laboratory of Neuroregeneration, Nantong University, Nantong 226007 PR China.
authors.forename: Yingjie
authors.initials: Y
authors.lastname: Wang
authors.forename: Yingying
authors.initials: Y
authors.lastname: Dong
authors.forename: Honghua
authors.initials: H
authors.lastname: Song
authors.forename: Yan
authors.initials: Y
authors.lastname: Liu
authors.forename: Mei
authors.initials: M
authors.lastname: Liu
authors.forename: Ying
authors.initials: Y
authors.lastname: Yuan
authors.forename: Fei
authors.initials: F
authors.lastname: Ding
authors.forename: Xiaosong
authors.initials: X
authors.lastname: Gu
authors.forename: Yongjun
authors.initials: Y
authors.lastname: Wang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D048430:Cell Shape; D003001:Cloning, Molecular; D017124:Conserved Sequence; D015870:Gene Expression; D055785:Gene Knockdown Techniques; D006801:Humans; D008116:Lizards; D008969:Molecular Sequence Data; D009416:Nerve Regeneration; D016501:Neurites / Q000378:metabolism / Q000502:physiology*; D009928:Organ Specificity; D010766:Phosphorylation; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D034741:RNA, Small Interfering / Q000235:genetics; D030162:Reptilian Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D050600:SNARE Proteins / Q000378:metabolism; D013116:Spinal Cord / Q000378:metabolism / Q000502:physiology*; D013572:Synaptic Vesicles / Q000378:metabolism; D050825:Synaptosomal-Associated Protein 25 / Q000235:genetics / Q000378:metabolism / Q000502:physiology*
chemical_list: D034741:RNA, Small Interfering; D030162:Reptilian Proteins; D050600:SNARE Proteins; D050825:Synaptosomal-Associated Protein 25