A phosphorylation epitope on MAP 1B that is transiently expressed in growing axons in the developing rat nervous system.
We have isolated a monoclonal antibody (150) that recognizes a phosphorylation epitope on the microtubule-associated protein (MAP) 1B. Immunoblot analysis of the developing rat central nervous system shows that monoclonal antibody 150 is directed against a protein of approximately 325 kDa (MAP 1B) that copolymerizes with microtubules through successive cycles of temperature-dependent assembly and disassembly. Furthermore, immunoprecipitated MAP 1B contains the epitope recognized by monoclonal antibody 150. Removal of phosphate from blotted proteins using alkaline phosphatase abolishes the binding of monoclonal antibody 150 to MAP 1B, indicating that the epitope is phosphorylated. In the developing rat nervous system, immunohistochemistry with monoclonal antibody 150 shows that the phosphorylation epitope on MAP 1B is transiently expressed in growing axons but not in dendrites. For instance, in the neonatal rat cerebellum, the parallel fibres of granule cells are stained only during elongation and not after synaptogenesis. The monoclonal antibody 150 epitope is also transiently expressed in radial glial fibres and in certain cell nuclei. All immunostaining of sections with monoclonal antibody 150 was completely abolished by alkaline phosphatase treatment. These observations and previous ones made by us in cell culture (Mansfield et al., J. Neurocytol., 20, 654-666, 1991) suggest that the phosphorylation epitope on MAP 1B recognized by monoclonal antibody 150, which has not been previously detected in vivo, may be important in axonogenesis.
Gordon-Weeks
P R
PR
Developmental Biology Research Centre, King's College London, UK.
Mansfield
S G
SG
Alberto
C
C
Johnstone
M
M
Moya
F
F
D000375:Aging / Q000378:metabolism*; D000818:Animals; D000911:Antibodies, Monoclonal; D001369:Axons / Q000502:physiology / Q000648:ultrastructure*; D001921:Brain / Q000166:cytology* / Q000254:growth & development; D002531:Cerebellum / Q000166:cytology* / Q000254:growth & development; D000939:Epitopes / Q000032:analysis*; D015151:Immunoblotting; D007150:Immunohistochemistry; D051379:Mice; D008807:Mice, Inbred BALB C / Q000276:immunology; D008869:Microtubule-Associated Proteins / Q000032:analysis / Q000096:biosynthesis*; D008870:Microtubules / Q000502:physiology / Q000648:ultrastructure; D010766:Phosphorylation; D051381:Rats
D000911:Antibodies, Monoclonal; D000939:Epitopes; D008869:Microtubule-Associated Proteins