title: Expression of nigrostriatal alpha 6-containing nicotinic acetylcholine receptors is selectively reduced, but not eliminated, by beta 3 subunit gene deletion.
abstract: mRNAs for the neuronal nicotinic acetylcholine receptor (nAChR) alpha6 and beta3 subunits are abundantly expressed and colocalized in dopaminergic cells of the substantia nigra and ventral tegmental area. Studies using subunit-null mutant mice have shown that alpha6- or beta3-dependent nAChRs bind alpha-conotoxin MII (alpha-CtxMII) with high affinity and modulate striatal dopamine release. This study explores the effects of beta3 subunit-null mutation on striatal and midbrain nAChR expression, composition, and pharmacology. Ligand binding and immunoprecipitation experiments using subunit-specific antibodies indicated that beta3-null mutation selectively reduced striatal alpha6* nAChR expression by 76% versus beta3(+/+) control. Parallel experiments showed a smaller reduction in both midbrain alpha3* and alpha6* nAChRs (34 and 42% versus beta3(+/+) control, respectively). Sedimentation coefficient determinations indicated that residual alpha6* nAChRs in beta3(-/-) striatum were pentameric, like their wild-type counterparts. Immunoprecipitation experiments on immunopurified beta3* nAChRs demonstrated that almost all wild-type striatal beta3* nAChRs also contain alpha4, alpha6, and beta2 subunits, although a small population of non-beta3 alpha6* nAChRs is also expressed. beta3 subunit incorporation seemed to increase alpha4 participation in alpha6beta2* complexes. (125)I-Epibatidine competition binding studies showed that the alpha-CtxMII affinity of alpha6* nAChRs from the striata of beta3(-/-) mice was similar to those isolated from beta3(+/+) animals. Together, the results of these experiments show that the beta3 subunit is important for the correct assembly, stability and/or transport of alpha6* nAChRs in dopaminergic neurons and influences their subunit composition. However, beta3 subunit expression is not essential for the expression of alpha6*, high-affinity alpha-CtxMII binding nAChRs.
authors.affiliation: Consiglio Nazionale delle Ricerche, Institute of Neuroscience, Section of Cellular and Molecular Pharmacology Center, Department of Medical Pharmacology, University of Milan, Via Vanvitelli 32, 20129 Milan, Italy. c.gotti@in.cnr.it
authors.forename: Cecilia
authors.initials: C
authors.lastname: Gotti
authors.forename: Milena
authors.initials: M
authors.lastname: Moretti
authors.forename: Francesco
authors.initials: F
authors.lastname: Clementi
authors.forename: Loredana
authors.initials: L
authors.lastname: Riganti
authors.forename: J Michael
authors.initials: JM
authors.lastname: McIntosh
authors.forename: Allan C
authors.initials: AC
authors.lastname: Collins
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Marks
authors.forename: Paul
authors.initials: P
authors.lastname: Whiteaker
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D003342:Corpus Striatum / Q000378:metabolism*; D017353:Gene Deletion; D005786:Gene Expression Regulation / Q000502:physiology; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008969:Molecular Sequence Data; D021122:Protein Subunits / Q000172:deficiency* / Q000235:genetics; D051381:Rats; D011978:Receptors, Nicotinic / Q000096:biosynthesis* / Q000172:deficiency* / Q000235:genetics; D013378:Substantia Nigra / Q000378:metabolism*
chemical_list: D021122:Protein Subunits; D011978:Receptors, Nicotinic; C104518:nicotinic receptor alpha6