Class III β-Tubulin isotype (β III) in the adrenal medulla: III. Differential expression of neuronal and glial antigens identifies two distinct populations of neuronal and glial-like (sustentacular) cells in the PC12 rat pheochromocytoma cell line maintained in a gelfoam matrix system
✍ Scribed by Katsetos, Christos D. ;Herman, Mary M. ;Balin, Brian J. ;Vinores, Stanley A. ;Hessler, Richard B. ;Arking, E. James ;Karkavelas, George ;Frankfurter, Anthony
- Publisher
- John Wiley and Sons
- Year
- 1998
- Tongue
- English
- Weight
- 755 KB
- Volume
- 250
- Category
- Article
- ISSN
- 0003-276X
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✦ Synopsis
Background:
The rat PC12 pheochromocytoma cell line provides an established system for the study of neuronal differentiation. To our knowledge, glial differentiation has not been reported in this cell line.
Methods: We have studied, by immunohistochemistry and immunoblotting, the presence of neuronal cytoskeletal antigens [class III -tubulin isotype ( III), microtubule associated proteins MAP2, MAP1B and tau, and different neurofilament (NF) protein components], and synaptophysin in comparison with the glial fibrillary acidic protein (GFAP) and S-100 protein in the PC12 cell line. In three different experiments, PC12 cells were maintained in a three-dimensional gelatin foam (Gelfoam) matrix system for up to 34 days with and without treatment with 1 mM dibutyryl cyclic (dc)AMP. Immunohistochemistry was performed on explants ranging from 2 to 32 days-in vitro, which were fixed in either Bouin's solution, 70% ethanol, or 10% neutral-buffered formalin and embedded in paraffin. Immunoblotting was performed on Gelfoam explants with a panel of antibodies against all aforementioned neuronal and glial markers. Additional immunoblot experiments using anti-GFAP and anti- III monoclonal antibodies in cell suspensions and homogenates from PC12 monolayer cultures were carried out to compare growth conditions in relation to the expression of these proteins.