effects of NGF and dibutyryl cAMP on neuronal differentiation of embryonal carcinoma cells |
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Authors: | Shanti Sharma John T. Hansen Mary F.D. Notter |
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Affiliation: | Department of Neurobiology and Anatomy, University of Rochester School of Medicine, NY 14642. |
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Abstract: | The P19S18O1A1 embryonal carcinoma cell line is capable of neuronal differentiation and is therefore useful in studying neuronal development and the influence of growth modulators on neuronal differentiation. We report here on the effects of nerve growth factor (NGF) and dibutyryl cyclic adenosine monophosphate (db cAMP), individually and combined, on differentiation of P19S18O1A1 cells. NGF alone did not induce any significant neuron-like changes in cultures exposed to NGF for as long as 12 days. Treatment with db cAMP resulted in changes in a significant population of the cells, including development of a neuron-like morphology, seen at both the light and electron microscopic level, loss of stage-specific embryonic antigen expression and the appearance of two neuronal markers, neurofilament protein and neuron-specific enolase. These changes were similar to changes seen when embryonal carcinoma (EC) cells are treated with retinoic acid. NGF in combination with dibutyryl cyclic adenosine monophosphate brought about similar changes as dibutyryl cyclic adenosine monophosphate alone, and was therefore not synergistic for induction of neuronal properties. In retinoic acid-treated cultures, the neuron-like cells had ultrastructural features very similar to neurons in non-tumorous, normal tissue, with typical organelles, such as one nucleolus, neurotubules and neurofilaments, while db cAMP-treated EC cells showed similar findings at the electronmicroscopic level. The results suggest that db cAMP can induce the neuronal phenotype in EC cells alone without pre-treatment with retinoic acid. |
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Keywords: | growth factors differentiation cell line neuronal phenotype neuronal proteins immunocytochemistry electronmicroscopy |
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