Characterization of functional nerve growth factor-receptors in a CNS glial cell line: monoclonal antibody 217c recognizes the nerve growth factor-receptor on C6 glioma cells.
The biological effects of nerve growth factor (NGF) have been shown to be mediated by the high-affinity form of the nerve growth factor receptor (NGF-R) in sympathetic and sensory neurons, and in PC12 cells. We report here that the central nervous system C6 rat glioma cell line likewise expresses functional high-affinity NGF-Rs. The expression of NGF-R mRNA in C6 cells can be up-regulated by cycloheximide and its own ligand, NGF; and it can be rapidly down-regulated by epidermal growth factor (EGF). Furthermore, C6 cells display NGF responsiveness by expressing c-fos mRNA within 30 minutes of treatment with NGF; and after 4-5 days of NGF exposure, C6 cells cease dividing as measured by [3H]-thymidine uptake, change shape, and reveal neurite-like processes. Scatchard analysis of [125I]-labelled NGF bound to solubilized C6 cells confirms the presence of both high- and low-affinity receptor protein. Crosslinking radiolabeled NGF to its receptor in the presence or absence of excess unlabeled NGF, followed by immunoprecipitation with monoclonal antibody (mAb) 192-IgG (a known anti-NGF-R antibody) and SDS-PAGE reveals a 100 kD band corresponding to the NGF/NGF-R complex. An identical band is observed when the immunoprecipitation is carried out with mAb 217c, suggesting that the 217c epitope is related to NGF-R. The 217c antibody was generated against C6 cells and shown to be a cell surface antibody (Peng et al., Science 215:1102-4, 1982); several investigators have used it subsequently as an immunocytochemical marker for Schwann cells. The significance of NGF-Rs in a CNS glial cell line is unclear, but association of NGF with the control of proliferation and/or differentiation of primitive glial cells is suggested.
Kumar
S
S
Laboratory of Biomedical and Environmental Sciences, University of California, Los Angeles 90024-1786.
Huber
J
J
Peña
L A
LA
Perez-Polo
J R
JR
Werrbach-Perez
K
K
de Vellis
J
J
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D004247:DNA / Q000235:genetics; D005910:Glioma / Q000473:pathology*; D009363:Neoplasm Proteins / Q000032:analysis; D009414:Nerve Growth Factors / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000032:analysis; D016760:Proto-Oncogene Proteins c-fos; D051381:Rats; D011956:Receptors, Cell Surface / Q000032:analysis* / Q000276:immunology; D017475:Receptors, Nerve Growth Factor; D014407:Tumor Cells, Cultured / Q000737:chemistry
D000911:Antibodies, Monoclonal; D009363:Neoplasm Proteins; D009414:Nerve Growth Factors; D011518:Proto-Oncogene Proteins; D016760:Proto-Oncogene Proteins c-fos; D011956:Receptors, Cell Surface; D017475:Receptors, Nerve Growth Factor; D004247:DNA