Nicotinic acetylcholine receptors (nAChRs) are an important class of ligand-gated ion channels that express throughout the central and peripheral nervous systems.  These receptors are composed of alpha (a1-a10) and beta (�2-�4) subunits as pentameric structures, and the correct subunit assembly and stoichiometry is essential for proper subcellular localization, agonist sensitivity, and ion channel Ca2+ permeability.  We examined spatially resolved individual receptors on the plasma membrane of living cells by exploiting the optical and spatial features of sub-wavelength nanostructures.  Zero-mode waveguides (diameter = 140 nm) were used to sequester small segments of the plasma membrane containing an individual receptor allowing us to overcome common issues associated with receptor diffusion and aggregation.  This approach allowed us to determine the stoichiometry of heteromeric nAChRs through single-molecule measurements at physiological densities of receptors ~500 times greater than accessible with previous optical techniques.

Single-molecule imaging of nicotinic receptors in live cells with zero-mode waveguide nanostructures

243rd National Meeting (2012)

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