title: GABAρ subunits confer a bicuculline-insensitive component to GFAP+ cells of cerebellum.
abstract: GABA-A receptors mediating synaptic or extrasynaptic transmission are molecularly and functionally distinct, and glial cells are known to express a plethora of GABA-A subunits. Here we demonstrate that GFAP(+) cells of the granular layer of cerebellum express GABAρ subunits during early postnatal development, thereby conferring peculiar pharmacologic characteristics to GABA responses. Electron microscopy revealed the presence of GABAρ in the plasma membrane of GFAP(+) cells. In contrast, expression in the adult was restricted to Purkinje neurons and a subset of ependymal cells. Electrophysiological studies in vitro revealed that astrocytes express functional receptors with an EC50 of 52.2 ± 11.8 μM for GABA. The evoked currents were inhibited by bicuculline (100 μM) and TPMPA (IC50, 5.9 ± 0.6 μM), indicating the presence of a GABAρ component. Coimmunoprecipitation demonstrated protein-protein interactions between GABAρ1 and GABAα1, and double immunofluorescence showed that these subunits colocalize in the plasma membrane. Three populations of GABA-A receptors in astrocytes were identified: classic GABA-A, bicuculline-insensitive GABAρ, and GABA-A-GABAρ hybrids. Clusters of GABA-A receptors were distributed in the perinuclear space and along the processes of GFAP(+) cells. Time-lapse microscopy showed GABAρ2-GFP accumulation in clusters located in the soma and along the processes. The clusters were relatively immobile, with mean displacement of 9.4 ± 0.9 μm and a net distance traveled of 1-2 μm, owing mainly to directional movement or simple diffusion. Modulation of GABAρ dynamics may be a novel mechanism of extrasynaptic transmission regulating GABAergic control of GFAP(+) cells during early postnatal development.
authors.affiliation: Departamento de Neurobiología Celular y Molecular, Laboratorio de Neurobiología Molecular y Celular, Instituto de Neurobiología, Universidad Nacional Autónoma de México, Juriquilla, 76230 Santiago de Querétaro, Querétaro, México.
authors.forename: Adriana
authors.initials: A
authors.lastname: Pétriz
authors.affiliation: Departamento de Neurobiología Celular y Molecular, Laboratorio de Neurobiología Molecular y Celular, Instituto de Neurobiología, Universidad Nacional Autónoma de México, Juriquilla, 76230 Santiago de Querétaro, Querétaro, México.
authors.forename: Daniel
authors.initials: D
authors.lastname: Reyes-Haro
authors.affiliation: Departamento de Neurobiología Celular y Molecular, Laboratorio de Neurobiología Molecular y Celular, Instituto de Neurobiología, Universidad Nacional Autónoma de México, Juriquilla, 76230 Santiago de Querétaro, Querétaro, México.
authors.forename: María Alejandra
authors.initials: MA
authors.lastname: González-González
authors.affiliation: Departamento de Neurobiología Celular y Molecular, Laboratorio de Neurobiología Molecular y Celular, Instituto de Neurobiología, Universidad Nacional Autónoma de México, Juriquilla, 76230 Santiago de Querétaro, Querétaro, México rmiledi@uci.edu ataulfo@inb.unam.mx.
authors.forename: Ricardo
authors.initials: R
authors.lastname: Miledi
authors.affiliation: Departamento de Neurobiología Celular y Molecular, Laboratorio de Neurobiología Molecular y Celular, Instituto de Neurobiología, Universidad Nacional Autónoma de México, Juriquilla, 76230 Santiago de Querétaro, Querétaro, México rmiledi@uci.edu ataulfo@inb.unam.mx.
authors.forename: Ataúlfo
authors.initials: A
authors.lastname: Martínez-Torres
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000166:cytology; D001640:Bicuculline / Q000737:chemistry*; D019070:Cell Lineage; D002462:Cell Membrane / Q000378:metabolism*; D002465:Cell Movement; D049109:Cell Proliferation; D002531:Cerebellum / Q000378:metabolism*; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D005904:Glial Fibrillary Acidic Protein / Q000737:chemistry*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D020128:Inhibitory Concentration 50; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D008856:Microscopy, Fluorescence; D009457:Neuroglia / Q000378:metabolism; D009474:Neurons / Q000378:metabolism; D018377:Neurotransmitter Agents / Q000378:metabolism*; D010721:Phosphinic Acids / Q000737:chemistry; D025941:Protein Interaction Mapping; D021381:Protein Transport; D011689:Purkinje Cells / Q000378:metabolism; D011725:Pyridines / Q000737:chemistry; D011963:Receptors, GABA-A / Q000502:physiology*; D013997:Time Factors
chemical_list: C101879:(1,2,5,6-tetrahydropyridin-4-yl)methylphosphinic acid; C547869:Gabrr1 protein, mouse; D005904:Glial Fibrillary Acidic Protein; D018377:Neurotransmitter Agents; D010721:Phosphinic Acids; D011725:Pyridines; D011963:Receptors, GABA-A; D049452:Green Fluorescent Proteins; D001640:Bicuculline
keywords: GABA-A receptor; GABAρ receptor; astrocytes; cerebellum; protein trafficking