Acquisition of neuronal proteins during differentiation of NG108-15 cells

The differentiated type of neuroblastoma x glioma hybrid cell line, NG108-1 5, has widely been used in in vitro studies instead of primary-cultured neu rons. Here we examined whether NG108-15 cells can be used as a model for st udying the neuronal differentiation process. We compared the expression of neuronal proteins (neurofilament 200 (NF200), phosphorylated-NF200 (p-NF200 ), microtubule associated protein 2, synaptophysin, syntaxin 1, choline ace tyltransferase, and acetylcholinesterase (AChE)) and a glial protein (vimen tin) between undifferentiated and differentiated NG108-15 cells by immunocy tochemistry and immunoblot analysis. The expression of all neuronal protein s, with the exception of NF200 and p-NF200, was positive in differentiated cells, but almost negative in undifferentiated cells. On the other hand, cy toskeletal intermediate filaments (NF200 and p-NF200) for neurons and that (vimentin) for glia were present in both undifferentiated and differentiate d cells. Furthermore, a high expression of AChE mRNA was confirmed in diffe rentiated cells by reverse transcription-PCR analysis. Our results showed t hat even though the expression of cytoskeletal filaments does not change du ring differentiation of NG108-15 cells, these cells during differentiation can serve as an appropriate tool for investigating and understanding the me chanisms involved in neuronal development and differentiation. (C) 2000 Els evier Science Ireland Ltd and the Japanese Neuroscience Society. All rights reserved.