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Nicotinic Acetylcholine Receptor Subtypes Expression during Rat Retina Development and Their Regulation by Visual Experience

CNR, Institute of Neuroscience, Cellular and Molecular Pharmacology, Department of Medical Pharmacology, and Center of Excellence on Neurodegenerative Diseases, University of Milan, Milan, Italy (M.M., S.V., L.R., F.C., C.G.); Department of Biomedical Sciences, Section of Physiology, University of Modena and Reggio Emilia, Modena, Italy (M.Z., G.L.); Consiglio Nazionale delle Ricerche, Institute of Chemistry of Molecular Recognition, Milan, Italy (R.L.); and Scuola Normale Superiore, Pisa, Italy (A.V.)

By acting through retinal nicotinic acetylcholine receptors (nAChRs), acetylcholine plays an important role in the development of both the retina and central visual pathways. Ligand binding and immunoprecipitation studies with subunit-specific antibodies showed that the expression of Bungarotoxin (Bgtx) and high-affinity epibatidine (Epi) receptors is regulated developmentally and increases until postnatal day 21 (P21). The increase in Epi receptors is caused by a selective increase in the subtypes containing the 2, 4, 6, 2, and 3 subunits. Immunopurification studies revealed three major populations of Epi receptors on P21: 6^* receptors (26%), which contain the 632, 6432, and 63/232 subtypes; 4(non-6)^* receptors (60%), which contain the 242 and 42 subtypes; and (non-4/non-6)^* receptors (14%), which contain the 22/4 and 32/4 subtypes. These three populations can be pharmacologically discriminated using conotoxin MII, which binds the 6^* population with high affinity. In situ hybridization showed that the transcripts for all of the subunits are heterogeneously distributed throughout retinal neurons at P21, with 3, 6, and 3 transcripts preferentially concentrated in the ganglion cell layer, 5 in the inner nuclear layer, and 4 and 2 distributed rather homogeneously. To investigate whether nAChR expression is affected by visual experience, we also studied dark-reared P21 rats. Visual deprivation had no effect on the expression of Bgtx receptors or the developmentally regulated Epi receptors containing the 2, 6, and/or 3 subunits but significantly increased the expression of the Epi receptors containing the 4 and 2 subunits. Overall, this study demonstrates that the retina is the rat neural region that expresses the widest array of nAChR subtypes. These receptors have a specific distribution, and their expression is finely regulated during development and by visual experience.

Address correspondence to: Dr. Cecilia Gotti, CNR, Institute of Neuroscience, Section of Cellular and Molecular Pharmacology, Department of Medical Pharmacology, University of Milan, Via Vanvitelli 32, 20129 Milano, Italy. E-mail: c.gotti{at}in.cnr.it

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