title: Two monoclonal antibodies recognising aa 634-668 and aa 1026-1055 of NogoA enhance axon extension and branching in cultured neurons.
abstract: In a previous study, we generated two monoclonal antibodies (mAbs) in mice, aNogoA-N and aNogo-66 mAb, which were raised against recombinant N-terminal fragments of rat NogoA and Nogo-66, respectively. When compared with the commercial rabbit anti-rat NogoA polyclonal antibody (pAb), which can specifically recognise NogoA, the two mAbs were also specific for the NogoA antigen in immunofluorescence histochemical (IHC) staining and Western blot (WB) analysis. Serial truncations of NogoA covering the N-terminal region of NogoA (aa 570-691) and Nogo-66 (aa 1026-1091) were expressed in E. coli. The epitopes recognised by aNogoA-N and aNogo-66 are located in the aa 634-668 and aa 1026-1055 regions of NogoA, respectively. Both mAbs remarkably enhanced the axon growth and branching of cultured hippocampal neurons in vitro. These results suggest that the antibodies that bind to aa 634-668 and aa 1026-1055 of NogoA may have stimulatory effects on axon growth and branching. Additionally, the two mAbs that we generated are specific for NogoA and significantly block NogoA function. In conclusion, two sites in NogoA located within aa 634-668 and aa 1026-1055 are recognised by our two antibodies and are novel and potentially promising targets for repair after central nervous system (CNS) injury.
authors.affiliation: Institute of Neurosciences, the Fourth Military Medical University, Xi'an, China ; Department of Anesthesiology, Stomatological College, Fourth Military Medical University, Xi'an, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Deng
authors.affiliation: Department of Clinical Laboratory, No. 174 Hospital of People's Liberation Army, Xiamen, China.
authors.forename: Fei
authors.initials: F
authors.lastname: Gao
authors.affiliation: Institute of Neurosciences, the Fourth Military Medical University, Xi'an, China.
authors.forename: Fang-Fang
authors.initials: FF
authors.lastname: Liu
authors.affiliation: Institute of Neurosciences, the Fourth Military Medical University, Xi'an, China.
authors.forename: Xiang-Hui
authors.initials: XH
authors.lastname: Zhao
authors.affiliation: Institute of Neurosciences, the Fourth Military Medical University, Xi'an, China.
authors.forename: Cai-Yong
authors.initials: CY
authors.lastname: Yu
authors.affiliation: Institute of Neurosciences, the Fourth Military Medical University, Xi'an, China.
authors.forename: Gong
authors.initials: G
authors.lastname: Ju
authors.affiliation: Department of Anesthesiology, Stomatological College, Fourth Military Medical University, Xi'an, China.
authors.forename: Li-Xian
authors.initials: LX
authors.lastname: Xu
authors.affiliation: Institute of Neurosciences, the Fourth Military Medical University, Xi'an, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Wang
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D001369:Axons / Q000502:physiology*; D002490:Central Nervous System / Q000293:injuries; D018604:Epitope Mapping; D000939:Epitopes / Q000737:chemistry; D019922:GAP-43 Protein / Q000737:chemistry; D006624:Hippocampus / Q000166:cytology; D007150:Immunohistochemistry; D008297:Male; D008856:Microscopy, Fluorescence; D009185:Myelin Proteins / Q000737:chemistry*; D009186:Myelin Sheath / Q000737:chemistry; D009474:Neurons / Q000166:cytology*; D000070798:Nogo Proteins; D017434:Protein Structure, Tertiary; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000737:chemistry
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D019922:GAP-43 Protein; D009185:Myelin Proteins; D000070798:Nogo Proteins; D011994:Recombinant Proteins; C000603201:Rtn4 protein, mouse; C000603197:Rtn4 protein, rat