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The long internal loop of the α3 subunit targets nAChRs to subdomains within individual synapses on neurons in vivo

Nature Neuroscience volume 1, pages 557–562 (1998)Cite this article

Abstract

Different types of neurotransmitter receptors coexist within single neurons and must be targeted to discrete synaptic regions for proper function. In chick ciliary ganglion neurons, nicotinic acetylcholine receptors (nAChRs) containing α3 and α5 subunits are concentrated in the postsynaptic membrane, whereas α-bungarotoxin receptors composed of α7 subunits are localized perisynaptically and excluded from the synapse. Using retroviral vector-mediated gene transfer in vivo, we show that the long cytoplasmic loop of α3 targets chimeric α7 subunits to the synapse and reduces endogenous nAChR surface levels, whereas the α5 loop does neither. These results show that a particular domain of one subunit targets specific receptor subtypes to the interneuronal synapse in vivo. Moreover, our findings suggest a difference in the mechanisms that govern assembly of interneuronal synapses as compared to the neuromuscular junction in vertebrates.

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Figure 1: Retroviral vector-mediated transfer of exogenous genes into CG neurons during synapse formation in vivo.
Figure 2: Electron micrographs demonstrating the distribution of overexpressed chimeric and wild-type α7 subunits in relation to synaptic and perisynaptic regions on CG neurons.
Figure 3: Schematic representation of the ultrastructural localization of endogenous nAChRs Bgt-nAChRs and chimeric subunits in synaptic and perisynaptic regions of ciliary ganglion neurons in vivo.
Figure 4: Surface levels of exogenous myc-tagged subunits and endogenous nAChRs on CG neurons.
Figure 5: Functional properties and surface levels of myc-tagged chimeric and wild-type α7 subunits expressed in Xenopus oocytes.

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Acknowledgements

This work is dedicated to Annette Altman. We acknowledge Donna Fekete and members of her lab for providing retroviral vector reagents, advice and assistance, Marc Ballivet for providing nAChR subunit clones and discussions, Josee Huard and Brian Moquin for their participation in early stages of these experiments, and Kathleen Dunlap, F.Rob Jackson and Stephen Lambert for comments on the manuscript. This work was supported by the Swiss National Science Foundation and OFES to D.B and NIH grant 21725 to M.H.J.

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Author notes

  1. Marjory Schwartz Levey

    Present address: Department of Biological Science, Wellesley College, 106 Central St., Wellesley, Massachusetts, 02181, USA

Authors and Affiliations

  1. Department of Neuroscience, Tufts University, School of Medicine, 136 Harrison Ave., Boston, 02111, Massachusetts, USA

    Brian M. Williams, Murali Krishna Temburni, Marjory Schwartz Levey & Michele H. Jacob

  2. Department of Physiology, University of Geneva, 1 Rue Michel-Servet, 4, 1211, Geneva, Switzerland

    Sonia Bertrand & Daniel Bertrand

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Correspondence to Michele H. Jacob.

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Williams, B., Temburni, M., Levey, M. et al. The long internal loop of the α3 subunit targets nAChRs to subdomains within individual synapses on neurons in vivo . Nat Neurosci 1, 557–562 (1998). https://doi.org/10.1038/2792

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