Neurofilament phosphorylation in axons and perikarya: immunofluorescence study of the rat spinal cord and dorsal root ganglia with monoclonal antibodies.
Rat dorsal root ganglia and spinal cord were stained with 12 monoclonal antibodies reacting with phosphorylated epitopes of two neurofilament proteins (NF 150K and NF 200K). Three monoclonal antibodies were axon-specific in both locations; neuronal perikarya were not stained. Nine monoclonal antibodies stained a subpopulation of neurofilament-positive sensory neurons, as indicated by double labeling experiments with polyclonal antibodies reacting with phosphorylated and dephosphorylated forms of the neurofilament protein triplet. Of these nine antibodies, two stained motor neuron perikarya in the spinal cord, while the remaining seven antibodies were axon-specific in this location. Subpopulations of stained and unstained motor neurons were not observed. With all 12 antibodies, the staining pattern in the lumbar dorsal root ganglia and spinal cord remained unchanged following sciatic nerve crush and ligature. The findings suggest that, in the neurofilament, some phosphorylated epitopes are axon specific, while other phosphorylated epitopes are present in both axons and perikarya. Furthermore, they suggest that differences exist between neuronal populations as to the presence of phosphorylated epitopes in perikaryal neurofilaments. It remains to be seen whether phosphorylation events in perikarya and axons have similar or different effects on neurofilament structure and function.
D000911:Antibodies, Monoclonal / Q000032:analysis; D001369:Axons / Q000032:analysis; D003599:Cytoskeleton / Q000032:analysis*; D005727:Ganglia, Spinal / Q000032:analysis* / Q000166:cytology; D007150:Immunohistochemistry; D007381:Intermediate Filament Proteins / Q000032:analysis*; D007382:Intermediate Filaments / Q000032:analysis*; D008970:Molecular Weight; D009046:Motor Neurons / Q000032:analysis; D016900:Neurofilament Proteins; D009475:Neurons, Afferent / Q000032:analysis; D010750:Phosphoproteins / Q000032:analysis*; D013116:Spinal Cord / Q000032:analysis* / Q000166:cytology
D000911:Antibodies, Monoclonal; D007381:Intermediate Filament Proteins; D016900:Neurofilament Proteins; D010750:Phosphoproteins