Single Particle Tracking of α7 Nicotinic AChR in Hippocampal Neurons Reveals Regulated Confinement at Glutamatergic and GABAergic Perisynaptic Sites

PLoS One

Christian Fuhrer¹  Jean-Marc Fritschy²  Thomas Bürli²  Corinne Sidler²  Helge Ewers³  Kristin Baer⁴ 

[1] Department of Neurochemistry, Brain Research Institute, University of Zurich, Zurich, Switzerland;Institute of Pharmacology and Toxicology, University of Zurich, Zurich, Switzerland;Laboratory of Physical Chemistry, ETH Zurich, Zurich, Switzerland;School of Medicine, Institute of Life Science, Swansea University, Swansea, United Kingdom

关键词: Interneurons;    Synapses;    Neurons;    Mass diffusivity;    Quantum dots;    Neuronal dendrites;    Ligand-gated ion channels;    Actins;   

DOI  :  10.1371/journal.pone.0011507

学科分类：医学（综合）

来源: Public Library of Science

【 摘 要 】

α7 neuronal nicotinic acetylcholine receptors (α7-nAChR) form Ca2+-permeable homopentameric channels modulating cortical network activity and cognitive processing. They are located pre- and postsynaptically and are highly abundant in hippocampal GABAergic interneurons. It is unclear how α7-nAChRs are positioned in specific membrane microdomains, particularly in cultured neurons which are devoid of cholinergic synapses. To address this issue, we monitored by single particle tracking the lateral mobility of individual α7-nAChRs labeled with α-bungarotoxin linked to quantum dots in live rat cultured hippocampal interneurons. Quantitative analysis revealed different modes of lateral diffusion of α7-nAChR dependent on their subcellular localization. Confined receptors were found in the immediate vicinity of glutamatergic and GABAergic postsynaptic densities, as well as in extrasynaptic clusters of α-bungarotoxin labeling on dendrites. α7-nAChRs avoided entering postsynaptic densities, but exhibited reduced mobility and long dwell times at perisynaptic locations, indicative of regulated confinement. Their diffusion coefficient was lower, on average, at glutamatergic than at GABAergic perisynaptic sites, suggesting differential, synapse-specific tethering mechanisms. Disruption of the cytoskeleton affected α7-nAChR mobility and cell surface expression, but not their ability to form clusters. Finally, using tetrodotoxin to silence network activity, as well as exposure to a selective α7-nAChR agonist or antagonist, we observed that α7-nAChRs cell surface dynamics is modulated by chronic changes in neuronal activity. Altogether, given their high Ca2+-permeability, our results suggest a possible role of α7-nAChR on interneurons for activating Ca2+-dependent signaling in the vicinity of GABAergic and glutamatergic synapses.

【 授权许可】

CC BY