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The Journal of Neuroscience, April 1, 2002, 22(7):2522-2529

Involvement of the 3 Subunit in Central Nicotinic Binding Populations

Paul Whiteaker¹, Cyrus G. Peterson¹, Wei Xu², J. Michael McIntosh³, Richard Paylor², Arthur L. Beaudet², Allan C. Collins¹, and Michael J. Marks¹

¹ Institute for Behavioral Genetics, University of Colorado, Boulder, Colorado 80309, ² Baylor College of Medicine, Department of Molecular and Human Genetics, Houston, Texas 77030, and ³ Departments of Biology and Psychiatry, University of Utah, Salt Lake City, Utah 84132

The 3 subunit gene was one of the first neuronal nicotinic acetylcholine receptor (nAChR) subunits to be cloned (Boulter et al., 1986), but direct evidence of 3 subunit contributions to mammalian central nAChR populations has not been presented. The studies reported here used mice engineered to contain a null mutation in the 3 nAChR subunit gene (Xu et al., 1999) to examine the involvement of the 3 subunit in central nAChR populations. Heterologously expressed 32 and 34 nAChRs are pharmacologically similar to native [¹²⁵I]-conotoxin MII (-CtxMII)-binding and 3-(2(S)-azetidinylmethoxy)pyridine dihydrochloride (A85380)-resistant [¹²⁵I]epibatidine-binding nAChR subtypes, respectively. The hypothesis that both native sites are 3-subtype nAChRs was tested using quantitative autoradiography in 3-null mutant mice. Somewhat surprisingly, deletion of the 3 nAChR subunit gene did not affect expression of the great majority of [¹²⁵I]-CtxMII-binding sites, indicating that they do not correspond to heterologously expressed 32 nAChRs. The only exception to this was observed in the habenulointerpeduncular tract, where 3-dependent [¹²⁵I]-CtxMII binding was observed. This finding may suggest the presence of an additional, minor nicotinic population in this pathway. In contrast, most A85380-resistant [¹²⁵I]epibatidine-binding nAChRs were dependent on 3 gene expression, suggesting that they do indeed correspond to an 3 nAChR subtype. However, widespread but lower levels of 3-independent A85380-resistant [¹²⁵I]epibatidine binding were also seen. Again, this may indicate the existence of an additional, minor population of non-3 A85380-resistant sites.

Key words: nicotinic acetylcholine receptor; 3 subunit; A85380-resistant binding; -conotoxin MII; autoradiography; 3 subunit-null mutant

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Copyright © 2002 Society for Neuroscience  0270-6474/02/2272522-08$05.00/0

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