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.
Highlights
After injury, the central nervous system (CNS) of adult mammals is limited in its ability to recover because of the inability of damaged axons to reconnect and regain their physiological structure and function [1]
The specificity and the affinity of the two monoclonal antibodies (mAbs) were tested by Western blot (WB) (Fig. 1A)
The two mAbs and the commercial rabbit antiNogoA polyclonal antibody bound to NogoA from spinal cord tissue, with corresponding bands at 200 kDa
Summary
The central nervous system (CNS) of adult mammals is limited in its ability to recover because of the inability of damaged axons to reconnect and regain their physiological structure and function [1]. In the past two decades, several CNS myelin-derived axon growth inhibitory factors have been found, including NogoA, myelin-associated glycoprotein (MAG), and oligodendrocyte myelin glycoprotein (OMgp) [4,5,6]. These proteins have been the subject of great research interest and are highly clinical relevant. A recent study reported that a region of NogoA (aa 290–562) attenuates cerebral ischaemia by inhibiting NADPH oxidasemediated oxidative damage and neuronal apoptosis [12], indicating that the functional diversity of different fragments of NogoA must be explored
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
