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Volume 16, Number 24, Issue of December 15, 1996 pp. 7892-7901
Copyright ©1996 Society for Neuroscience

Neuronal Nicotinic Receptor Expression in Sensory Neurons of the Rat Trigeminal Ganglion: Demonstration of alpha 3beta 4, a Novel Subtype in the Mammalian Nervous System

Received July 16, 1996; revised Sept. 25, 1996; accepted Oct. 1, 1996.

Christopher M. Flores1, Raquel M. DeCamp1, Sonja Kilo1, Scott W. Rogers2, and Kenneth M. Hargreaves1

1 Department of Restorative Sciences, University of Minnesota, Minneapolis, Minnesota 55455, and 2 Salt Lake City Veterans Administration-Geriatric Research Education Clinical Center and Department of Neurobiology and Anatomy, University of Utah, Salt Lake City, Utah 84112

The identification of a family of neuronal nicotinic receptor subunit genes establishes the potential for multiple subtypes with diverse physiological functions. Virtually all of the high affinity nicotinic receptors measured to date in the rodent CNS are composed of alpha 4 and beta 2 subunits only. However, the demonstration of other subunit transcripts in a variety of central and peripheral nervous tissues suggests a greater degree of receptor subtype heterogeneity than so far has been elucidated. The purpose of the present studies was to determine at the mRNA and protein levels which neuronal nicotinic receptor subunits are expressed by sensory neurons of the rat trigeminal ganglion and in what combinations these gene products associate to form neuronal nicotinic receptor subtypes in this tissue. Radioreceptor binding analysis indicated that in the adult rat trigeminal ganglion there exist at least two nicotinic receptor binding sites with differing affinities for [3H]-epibatidine. In situ hybridization histochemical studies revealed the existence of mRNA encoding the alpha 3, alpha 4, alpha 5, beta 2, and beta 4 subunits, but not the alpha 2 subunit. Immunoprecipitation with subunit-specific antisera demonstrated that each of the subunits present in the ganglion at the mRNA level is a constituent of nicotinic receptors capable of binding 3H-epibatidine. Various applications of these approaches yielded strong evidence that, in addition to alpha 4beta 2, which is thought to be the predominant neuronal nicotinic receptor subtype in the rodent CNS, trigeminal sensory neurons express as the principal subtype alpha 3beta 4, which has not been demonstrated previously in mammalian nervous tissue.

Key words: nicotinic receptor subtype; sensory neurons; trigeminal ganglion; radioreceptor binding; [3H]-epibatidine; immunoprecipitation; subunit composition; in situ hybridization; mRNA




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