Antigenic expression of neuron-associated class III beta-tubulin isotype (h beta 4) and microtubule-associated protein 2 (MAP2) by the human retinoblastoma cell line WERI-Rb1. A comparative immunoblot and immunocytochemical study.
The antigenic expression of two neuron-associated microtubule proteins, class III beta-tubulin isotype (h beta 4) and microtubule-associated protein 2 (MAP2), was evaluated in a comparative immunoblot and immunocytochemical study of the human retinoblastoma cell line WERI-Rb1 maintained for up to 30 days in three different in vitro conditions. Western blots were performed on whole sodium dodecyl sulfate extracts of cells grown in floating suspensions, on Gelfoam matrices and on coverslips. Immunoperoxidase histochemistry was performed on matrix cultures. Immunoblotting demonstrated that h beta 4 and MAP2 were present under all culture conditions. By immunocytochemistry, staining of cytologically undifferentiated cells with anti-h beta 4 and anti-MAP2 monoclonal antibodies was found on Gelfoam matrix explants. In contrast, glial fibrillary acidic protein was not detected by either immunoblots or immunocytochemistry. These findings are in keeping with the solely neuroblastic nature of this line and provide no evidence for its divergent (i.e. neuronal and glial) differentiation capacity.
Gass
P
P
Department of Pathology, University of Virginia School of Medicine, Charlottesville.
Frankfurter
A
A
Katsetos
C D
CD
Herman
M M
MM
Donoso
L A
LA
Rubinstein
L J
LJ
D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D005134:Eye Neoplasms / Q000378:metabolism; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D006801:Humans; D007124:Immunoenzyme Techniques; D008869:Microtubule-Associated Proteins / Q000096:biosynthesis*; D012160:Retina / Q000378:metabolism; D012175:Retinoblastoma / Q000378:metabolism; D014404:Tubulin / Q000378:metabolism*; D014407:Tumor Cells, Cultured / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D005904:Glial Fibrillary Acidic Protein; D008869:Microtubule-Associated Proteins; D014404:Tubulin