Absence of neuron-associated microtubule proteins in the rat C-6 glioma cell line. A comparative immunoblot and immunohistochemical study.
Three neuron-associated microtubule proteins, Class III beta-tubulin isotype, MAP-2, and tau, were evaluated in a comparative immunoblot and immunohistochemical study of the rat C-6 glioma cell line maintained for up to 31 days in vitro. Western blots on whole SDS extracts of cells grown: (i) as monolayers on plastic dishes (for 13 and 16 days); (ii) as monolayers on poly-D-lysine coated glass coverslips (for 3, 7, and 11 days); and (iii) as explants on Gelfoam matrices (for 10, 30, and 31 days) were probed with monoclonal antibodies (MoAb) specific for the above-mentioned microtubule proteins. For these and all other markers employed, immunoperoxidase histochemistry was performed only on the matrix cultures. The immunoblot experiments demonstrated that the Class III beta-tubulin isotype, MAP2, and tau were not expressed by the C-6 cell line in any of the culture conditions, nor were they found by immunohistochemistry. In contrast, explants from all culture conditions were positive for glial fibrillary acidic (GFA) protein and for a universal anti-beta-tubulin isotype MoAb by immunoblotting, as well as by immunohistochemistry in Gelfoam matrix cultures maintained in an organ culture system. Both sets of experiments indicate that these markers are not altered under three different conditions of growth over a one-month period in vitro. The expression of GFA protein and the absence of detectable levels of Class III beta-tubulin, MAP2, and tau are in keeping with the astrocytic phenotype of the C-6 cell line.
Zientek
G M
GM
Department of Pathology, (Neuropathology), University of Virginia School of Medicine, Charlottesville.
Herman
M M
MM
Katsetos
C D
CD
Frankfurter
A
A
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D015153:Blotting, Western; D001932:Brain Neoplasms / Q000378:metabolism*; D002454:Cell Differentiation; D005910:Glioma / Q000378:metabolism*; D007124:Immunoenzyme Techniques; D007150:Immunohistochemistry; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D009474:Neurons / Q000378:metabolism*; D051381:Rats; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D008869:Microtubule-Associated Proteins