Abstract
AbstractWe recently established adrenal medullary cell line tsAM5D, immortalized with a temperature‐sensitive mutant of the oncogene simian virus 40 large T‐antigen. When cultured with basic fibroblast growth factor (bFGF) and ciliary neurotrophic factor (CNTF), tsAM5D cells proliferated at the permissive temperature (33°C) for the expression of the oncogene and differentiated into neuron‐like cells at the nonpermissive temperature (39°C). In the present study, we investigated the gene expression of several neurotrophic factors associated with the neuronal differentiation induced by the temperature shift. Using real‐time RT‐PCR analysis on bFGF and CNTF‐treated cells, we found up‐regulated expression of mRNAs for bFGF, leukemia inhibitory factor, transforming growth factor (TGF)‐β2, persephin, acidic fibroblast growth factor, neurotrophin‐3, preproenkephalin, and TGF‐β1 and down‐regulated expression of the mRNAs for interleukin‐6, neurturin, and CNTF. No apparent differences were observed in the mRNA level of insulin‐like growth factor‐2, TGF‐β3, and neurotrophin‐4/5. These results suggest that the regulated expression of neurotrophic factor genes play an important role in the neuronal differentiation of tsAM5D cells.
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