Developmentally regulated masking of an intracellular epitope of the 180 kDa isoform of the neural cell adhesion molecule NCAM.
The neural cell adhesion molecule NCAM is a cell surface glycoprotein that occurs in several isoforms. It was previously shown that the largest 180-kDa isoform of NCAM (NCAM 180) accumulates at sites of cell contact and in postsynaptic densities and may be responsible for the stabilization of cell contacts by its interaction with the membrane-cytoskeleton linker protein brain spectrin. In immunohistochemical studies on the expression of the NCAM 180, we noticed that two NCAM 180 specific monoclonal antibodies, termed 481 and D3, showed different patterns of immunoreactivity in sections of fresh-frozen adult mouse brain. Here we show that the D3-specific, but not the 481-specific epitope becomes inaccessible to the antibody during development of the hippocampal formation, coincident with the establishment of stable cell-cell contacts. In contrast, in the olfactory bulb with its continually regenerating olfactory nerve fibers, both NCAM 180 antibodies remain fully immunoreactive throughout development and adulthood. We also show that the D3-specific epitope becomes inaccessible in primary cerebellar neuron cultures with time in culture. Electrophoretic separation of hippocampal membrane proteins under nondenaturating conditions showed NCAM to be present in protein complexes of different molecular weights at different developmental stages. We propose that NCAM is involved in the formation of developmentally regulated, noncovalent complexes with as yet unknown partner molecules that could be responsible for the masking of the D3-specific epitope.
Kramer
I
I
Department of Neurobiology, Swiss Federal Institute of Technology, Hoenggerberg, Zürich.
Hall
H
H
Bleistein
U
U
Schachner
M
M
D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D002454:Cell Differentiation / Q000276:immunology; D002478:Cells, Cultured; D002531:Cerebellum / Q000166:cytology / Q000254:growth & development / Q000276:immunology*; D000939:Epitopes; D006624:Hippocampus / Q000166:cytology / Q000276:immunology; D051379:Mice; D008813:Mice, Inbred ICR; D008815:Mice, Inbred Strains; D008970:Molecular Weight; D019006:Neural Cell Adhesion Molecules / Q000276:immunology*; D009474:Neurons / Q000166:cytology / Q000276:immunology*; D010446:Peptide Fragments / Q000276:immunology; D011994:Recombinant Proteins / Q000276:immunology
D000911:Antibodies, Monoclonal; D000939:Epitopes; D019006:Neural Cell Adhesion Molecules; D010446:Peptide Fragments; D011994:Recombinant Proteins