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Developmental Expression of Heteromeric Nicotinic Receptor Subtypes in Chick Retina

Consiglio Nazionale delle Ricerche (CNR), Institute of Neuroscience, Cellular and Molecular Pharmacology Section, Department of Medical Pharmacology and Center of Excellence on Neurodegenerative Diseases, University of Milan, Milan, Italy (S.V., M.M., F.C., C.G.); CNR Center of Hormone Chemistry, Milan, Italy (R.L.); and Laboratory of Molecular Pharmacology, Department of Pharmacological Sciences, University of Milan, Italy (G.E.R.)

Acting through nicotinic acetylcholine receptors (nAChRs), acetylcholine plays an important role in retinal development and the formation of retinal connections to target tissues, but very little is known about the nAChR subtypes expressed in vertebrate retina during neuronal development. We used immunoprecipitation and [³H]epibatidine binding to study the expression of chick retina -bungarotoxin–insensitive heteromeric nAChRs during development and adulthood, and found that it is strictly developmentally regulated, reaching a peak on postnatal day 1. The increase in [³H]epibatidine receptors is caused mainly by an increase in the receptors containing the 2, 6, 3, and 4 subunits. The contribution of subunits to [³H]epibatidine receptors significantly changes during development: the 2 subunit is contained in the majority (84%) of receptors on embryonic day (E) 7 but in only 32% on postnatal day (P) 1, whereas the 4-containing receptors increase from 22% to 78% during the same period. Using a sequential immunodepletion procedure, we purified the 2- and 4-containing subtypes and found that they coassemble with 4 and/or 3 on E11, and also with the 2, 6, and 3 on P1. After the immunodepletion of 6-containing receptors, the 2- and 4-containing receptors have a very similar pharmacological profile on P1. Parallel immunoprecipitation experiments in other brain areas showed that the developmentally regulated receptors in optic lobe are those containing the 2, 5, and 2 subunits and those containing the 4 and 2 subunits, whereas the receptors in forebrain-cerebellum contain the 4 and 2 subunits with or without the 5 subunit. These results indicate that there is an increase in receptor heterogeneity and complexity in chick retina during development that is also maintained in adulthood.

Received January 13, 2003; accepted February 12, 2003.

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}csfic.mi.cnr.it

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