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Volume 17, Number 2,
Issue of January 15, 1997
pp. 586-596
Copyright ©1997 Society for Neuroscience
Heterogeneity of Nicotinic Receptor Class and Subunit mRNA
Expression among Individual Parasympathetic Neurons from Rat
Intracardiac Ganglia
Received June 21, 1996; revised Oct. 22, 1996; accepted Oct. 24, 1996.
Kevin Poth,
Thomas J. Nutter,
Javier Cuevas,
Michael
J. Parker,
David J. Adams, and
Charles W. Luetje
Department of Molecular and Cellular Pharmacology, University of
Miami School of Medicine, Miami, Florida 33101
Neurons have the potential to form thousands of distinct neuronal
nicotinic receptors from the eight  and three  subunits that
currently are known. In an effort to determine how much of this
potential complexity is realized among individual neurons, we examined
the nicotinic pharmacological and biophysical properties and receptor
subunit mRNA expression patterns in individual neurons cultured from
rat epicardial ganglia. Analysis of the whole-cell pharmacology of
these neurons showed a diversity of responses to the agonists
acetylcholine, nicotine, cytisine, and
1,1-dimethyl-4-phenylpiperazinium, suggesting that a heterogeneous
population of nicotinic receptor classes, or subtypes, is expressed by
individual neurons. Single-channel analysis demonstrated three distinct
conductances (18, 24, and 31 pS), with patches from different neurons
containing different combinations of these channel classes. We used
single-cell RT-PCR to examine nicotinic acetylcholine receptor (nAChR)
subunit mRNA expression by individual neurons. Although mRNAs encoding
all eight neuronal nAChR subunits for which we probed (2-5,
7, 2-4) were present in multicellular cultures, we found that
individual epicardial neurons express distinct subsets of these nAChR
subunit mRNAs. These results suggest that individual epicardial neurons express distinct arrays of nAChR subunits and that these subunits may
assemble into functional receptors with distinct and variable subunit
composition. This variable receptor subunit expression provides an
explanation for the diversity of pharmacological and single-channel
responses we have observed in individual neurons.
Key words:
neuronal nicotinic receptors;
nicotinic acetylcholine
receptors;
cardiac ganglia;
parasympathetic ganglia;
single-cell
RT-PCR;
nicotinic pharmacology
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