Constitutive expression of the mature array of neurofilament proteins by a CNS neuronal cell line.
Neurofilament protein expression was examined immunochemically in a neuronal cell line derived from postnatal day 21 septal tissue. The SN48.1p cell line was found to constitutively synthesize an array of neurofilament proteins typical of a mature neuron. All three neurofilament subunits (NF-L, NF-M, and NF-H) as well as differentially phosphorylated isoforms (P-, P+, P++, and P ) of NF-M and NF-H were identified by immunoblot analysis. Immunofluorescence studies revealed that the neurofilament proteins were components of discrete, filamentous structures. Abnormal intracellular aggregations of neurofilament proteins were never observed. Some SN48.1p cells apportioned specific isoforms into selected intracellular regions based on the molecular weight and phosphorylation level of the protein. NF-L was preferentially localized to perikarya and proximal neurites; NF-M[P++] and NF-H[P ] were distributed to distal aspects of neurites. The expression of these differentiated features of neurofilament proteins and, presumably, the synthesis of the kinases and phosphatases required for normal neurofilament metabolism occurred in the absence of growth factors, differentiating agents, and specialized culture substrates. In addition, the non-neuronal intermediate filaments glial fibrillary acidic protein and epithelial cytokeratin proteins were absent. These data demonstrate that SN48.1p cells exhibit a neurofilament phenotype characteristic of mature neurons and provide a unique model to examine the expression and function of neurofilaments in differentiated neuronal cells.
Lee
H J
HJ
Committee on Neurobiology, University of Chicago, IL 60637.
Elliot
G J
GJ
Hammond
D N
DN
Lee
V M
VM
Wainer
B H
BH
D000818:Animals; D000906:Antibodies / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000276:immunology; D001921:Brain / Q000166:cytology / Q000378:metabolism; D001923:Brain Chemistry / Q000502:physiology; D002460:Cell Line; D002999:Clone Cells / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D019922:GAP-43 Protein; D005904:Glial Fibrillary Acidic Protein / Q000096:biosynthesis; D015151:Immunoblotting; D007527:Isoenzymes; D008562:Membrane Glycoproteins / Q000096:biosynthesis; D051379:Mice; D009419:Nerve Tissue Proteins / Q000096:biosynthesis; D016501:Neurites / Q000378:metabolism; D016900:Neurofilament Proteins / Q000032:analysis / Q000096:biosynthesis*; D009474:Neurons / Q000378:metabolism*; D010641:Phenotype; D014407:Tumor Cells, Cultured
D000906:Antibodies; D000911:Antibodies, Monoclonal; D019922:GAP-43 Protein; D005904:Glial Fibrillary Acidic Protein; D007527:Isoenzymes; D008562:Membrane Glycoproteins; D009419:Nerve Tissue Proteins; D016900:Neurofilament Proteins; C051774:neurofilament protein H; C061195:neurofilament protein M