Regulation of CNS glial phenotypes in N20.1 cells

The N20.1 oligodendroglial cell line, immortalized with SV40 T antigen, simultaneously expresses oligodendroglial markers and glial fibrillary acidic protein (GFAP), an astroglial marker. This study examines the plasticity of N20.1 cells with regard to GFAP expression, and its relationship to expression of SV40 T antigen, p53, and a novel nuclear antigen detected by the A007 monoclonal antibody. Marked changes occur in GFAP levels and cell morphology when N20.1 cells are switched from the permissive temperature (34degreesC) to the non-permissive temperature (39degreesC), and with cyclic AMP elevation at 39degreesC. At 34degreesC, levels of GFAP are high; when cells are switched to 39degreesC, GFAP levels decrease significantly, then increase slightly when forskolin is added. At both temperatures, the cells display feathery GFAP immunostaining. When forskolin is added at 39degreesC, however, cells display bright fibrous GFAP staining in elongated processes. The changes in GFAP were compared to changes in T antigen and p53. As expected, the decrease in T antigen at 39degreesC was accompanied by movement of p53 from the nucleus to cytoplasm. Total p53 levels did not change, however, and forskolin did not alter the respective distribution or levels of p53 at either temperature. At both temperatures, the cell bodies and processes show internal expression of sulfatide, as demonstrated with the O4, Sulph I, and A007 antibodies. We show, for the first time, abundant nuclear immunoreactivity with the A007 monoclonal antibody in the N20.1 cells. This nuclear reactivity is seen at 34degreesC, but not at 39degreesC, similar to p53, and is not detected with the other sulfatide antibodies. Double-label immunostaining shows that the nuclear A007 immunoreactivity is colocalized in nuclear structures with T antigen and p53 at 34degreesC, but is not found in every nucleus containing these antigens. We conclude that regulation of GFAP expression and morphology in N20.1 cells is dependent on a combination of T antigen expression and level of cAMP and may be related to regulation of p53 and the A007 nuclear antigen. (C) 2003 Wiley-Liss, Inc.