Differential expression of sodium channels and nicotinic acetylcholine receptor channels in nnr variants of the PC12 pheochromocytoma cell line.
An important component of neuronal differentiation is the tightly controlled expression of a spectrum of ion channel proteins. Ion channels play a critical role in the generation and propagation of action potentials as well as in the cellular response to neurotransmitters, and thus are central in the transfer and integration of information in the nervous system. A model system amenable to analysis of ion channel expression and neuronal differentiation is the rat pheochromocytoma (PC12) cell line. Here, we have used electrophysiological and molecular biological approaches to analyze the expression of voltage-dependent sodium (Na) channels and nicotinic acetylcholine receptors (nAChR) in mutagenized variants (nnr cells) of the PC12 cell line. Our data reveal striking differences in the expression of these channels when compared to wild-type PC12 cells. Even in the absence of nerve growth factor (NGF), nnr cells express functional Na channels and Na channel mRNA at levels exceeding those in wild-type PC12 cells differentiated with NGF. In contrast, acetylcholine-induced currents were evident in only a small proportion of cells, presumably due to the altered pattern of expression of mRNAs encoding individual nAChR subunits. The altered ion channel expression in these variants provides an avenue for analyzing Na channel and nAChR channel function, as well as for identifying mechanisms governing their expression.
Fanger
G R
GR
Department of Biochemistry, Dartmouth Medical School, Hanover, New Hampshire 03755-3833, USA.
Brennan
C
C
Henderson
L P
LP
Gardner
P D
PD
Maue
R A
RA
D000818:Animals; D002454:Cell Differentiation; D002999:Clone Cells; D015972:Gene Expression Regulation, Neoplastic; D016296:Mutagenesis; D009363:Neoplasm Proteins / Q000096:biosynthesis / Q000235:genetics; D009414:Nerve Growth Factors / Q000494:pharmacology; D009419:Nerve Tissue Proteins / Q000096:biosynthesis* / Q000235:genetics; D016716:PC12 Cells / Q000378:metabolism*; D018408:Patch-Clamp Techniques; D051381:Rats; D020917:Receptor, trkA / Q000378:metabolism; D011978:Receptors, Nicotinic / Q000096:biosynthesis* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D015222:Sodium Channels / Q000096:biosynthesis* / Q000235:genetics; D014162:Transfection
D009363:Neoplasm Proteins; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins; D011978:Receptors, Nicotinic; D011993:Recombinant Fusion Proteins; D015222:Sodium Channels; D020917:Receptor, trkA