PMID: 9443895Apr 4, 1998Paper

Contiguous phosphorylated and non-phosphorylated domains along axonal neurofilaments

Journal of Cell Science
Anthony Brown

Abstract

I have investigated the phosphorylation state of the medium molecular mass neurofilament protein (NF-M) along axonal neurofilaments. Cultured embryonic sensory neurons were treated with non-ionic detergent to cause the cytoskeletal polymers to splay apart from each other. Neurofilaments were visualized by double-label immunofluorescence microscopy and the proportion of their length that stained with various NF-M antibodies was determined using digital image analysis techniques. Monoclonal antibody RMO255, which binds to NF-M independently of phosphorylation state, stained an average of 98% of the neurofilament length. In contrast, monoclonal antibody RMO55, which binds specifically to a phosphorylated epitope on NF-M, stained some neurofilaments completely, some not at all, and some along part of their length. These partly stained neurofilaments exhibited single or multiple discrete segments of staining along their length separated by segments that were unstained. The average proportion of the neurofilament length that stained with this antibody was lowest proximally (12-22%, n=3) and increased along the axon to reach a maximum distally (58-87%, n=3). A converse pattern (77-87% proximally and 2-9% distally, n=3) was observed fo...Continue Reading

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