Acquisition of neuronal proteins during differentiation of NG108-15 cells.
The differentiated type of neuroblastomaxglioma hybrid cell line, NG108-15, has widely been used in in vitro studies instead of primary-cultured neurons. Here we examined whether NG108-15 cells can be used as a model for studying 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 acetyltransferase, and acetylcholinesterase (AChE)) and a glial protein (vimentin) between undifferentiated and differentiated NG108-15 cells by immunocytochemistry and immunoblot analysis. The expression of all neuronal proteins, with the exception of NF200 and p-NF200, was positive in differentiated cells, but almost negative in undifferentiated cells. On the other hand, cytoskeletal intermediate filaments (NF200 and p-NF200) for neurons and that (vimentin) for glia were present in both undifferentiated and differentiated cells. Furthermore, a high expression of AChE mRNA was confirmed in differentiated cells by reverse transcription-PCR analysis. Our results showed that even though the expression of cytoskeletal filaments does not change during differentiation of NG108-15 cells, these cells during differentiation can serve as an appropriate tool for investigating and understanding the mechanisms involved in neuronal development and differentiation.
Tojima
T
T
Division of Biological Sciences, Graduate School of Science, Hokkaido University, North 10, West 8, Kita-ku, 060-0810, Sapporo, Japan.
Yamane
Y
Y
Takahashi
M
M
Ito
E
E
D000110:Acetylcholinesterase / Q000235:genetics; D000818:Animals; D002454:Cell Differentiation / Q000502:physiology; D015151:Immunoblotting; D007150:Immunohistochemistry; D051379:Mice; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D009457:Neuroglia / Q000378:metabolism; D009474:Neurons / Q000166:cytology* / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014407:Tumor Cells, Cultured
D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; D000110:Acetylcholinesterase