title: Synaptic GABAA receptors are directly recruited from their extrasynaptic counterparts.
abstract: GABAA receptors mediate the majority of fast synaptic inhibition in the brain. The accumulation of these ligand-gated ion channels at synaptic sites is a prerequisite for neuronal inhibition, but the molecular mechanisms underlying this phenomenon remain obscure. To further understand these processes, we have examined the cellular origins of synaptic GABAA receptors. To do so, we have created fluorescent GABAA receptors that are capable of binding -bungarotoxin (Bgt), facilitating the visualization of receptor endocytosis, exocytosis and delivery to synaptic sites. Imaging with Bgt in hippocampal neurons revealed that GABAA receptor endocytosis occurred exclusively at extrasynaptic sites, consistent with the preferential colocalization of extrasynaptic receptors with the AP2 adaptin. Receptor insertion into the plasma membrane was also predominantly extrasynaptic, and pulse-chase analysis revealed that these newly inserted receptors were then able to access directly synaptic sites. Therefore, our results demonstrate that synaptic GABAA receptors are directly recruited from their extrasynaptic counterparts. Moreover, they illustrate a dynamic mechanism for neurons to modulate GABAA receptor number at inhibitory synapses by controlling the stability of extrasynaptic receptors.
authors.affiliation: Department of Neuroscience, University of Pennsylvania, PA 19104, USA.
authors.forename: Yury
authors.initials: Y
authors.lastname: Bogdanov
authors.forename: Guido
authors.initials: G
authors.lastname: Michels
authors.forename: Cecilia
authors.initials: C
authors.lastname: Armstrong-Gold
authors.forename: Philip G
authors.initials: PG
authors.lastname: Haydon
authors.forename: Jon
authors.initials: J
authors.lastname: Lindstrom
authors.forename: Menelas
authors.initials: M
authors.lastname: Pangalos
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Moss
mesh_terms: D000818:Animals; D001693:Biological Transport, Active; D002038:Bungarotoxins; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D002966:Clathrin / Q000378:metabolism; D004705:Endocytosis; D005456:Fluorescent Dyes; D006624:Hippocampus / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D009474:Neurons / Q000378:metabolism; D011963:Receptors, GABA-A / Q000235:genetics / Q000378:metabolism*; D011978:Receptors, Nicotinic / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013569:Synapses / Q000378:metabolism*; D064569:alpha7 Nicotinic Acetylcholine Receptor
chemical_list: D002038:Bungarotoxins; C579167:Chrna7 protein, human; D002966:Clathrin; D005456:Fluorescent Dyes; D011963:Receptors, GABA-A; D011978:Receptors, Nicotinic; D011993:Recombinant Fusion Proteins; D064569:alpha7 Nicotinic Acetylcholine Receptor