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      Immunohistochemistry of GluR1 subunits of AMPA receptors of rat cerebellar nerve cells

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          Abstract

          The localization of GluR1 subunits of ionotropic glutamate receptors in the glial cells and inhibitory neurons of cerebellar cortex and their association with the climbing and parallel fibers, and basket cell axons were studied. Samples of P14 and P21 rat cerebellar cortex were exposed to a specific antibody against GluR1 subunit(s) of AMPA receptors and were examined with confocal laser scanning microscopy. GluR1 strong immunoreactivity was confined to Purkinje cell and the molecular layer. Weak GluR1 immunoreactivity was observed surrounding some Golgi cells in the granule cell layer. Intense GluR1 immunoreactivity was localized around Purkinje, basket, and stellate cells. Purkinje cells expressed strong GluR1 immunoreactivity surrounding the cell body, primary dendritic trunk and secondary and tertiary spiny dendritic branches. Marked immunofluorescent staining was also detected in the Bergmann glial fibers at the level of middle and outer third molecular layer. Positive immunofluorescence staining was also observed surrounding basket and stellate cells, and in the capillary wall. These findings suggest the specific localization of GluR1 subunits of AMPA receptors in Bergmann glial cells, inhibitory cerebellar neurons, and the associated excitatory glutamatergic circuits formed by climbing and parallel fibers, and by the inhibitory basket cell axons.

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          Synaptic activity at calcium-permeable AMPA receptors induces a switch in receptor subtype.

          Activity-dependent change in the efficacy of transmission is a basic feature of many excitatory synapses in the central nervous system. The best understood postsynaptic modification involves a change in responsiveness of AMPAR (alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptor)-mediated currents following activation of NMDA (N-methyl-D-aspartate) receptors or Ca2+-permeable AMPARs. This process is thought to involve alteration in the number and phosphorylation state of postsynaptic AMPARs. Here we describe a new form of synaptic plasticity--a rapid and lasting change in the subunit composition and Ca2+ permeability of AMPARs at cerebellar stellate cell synapses following synaptic activity. AMPARs lacking the edited GluR2 subunit not only exhibit high Ca2+ permeability but also are blocked by intracellular polyamines. These properties have allowed us to follow directly the involvement of GluR2 subunits in synaptic transmission. Repetitive synaptic activation of Ca2+-permeable AMPARs causes a rapid reduction in Ca2+ permeability and a change in the amplitude of excitatory postsynaptic currents, owing to the incorporation of GluR2-containing AMPARs. Our experiments show that activity-induced Ca2+ influx through GluR2-lacking AMPARs controls the targeting of GluR2-containing AMPARs, implying the presence of a self-regulating mechanism.
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            Glia-synapse interaction through Ca2+-permeable AMPA receptors in Bergmann glia.

            Glial cells express a variety of neurotransmitter receptors. Notably, Bergmann glial cells in the cerebellum have Ca2+-permeable alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-type glutamate receptors (AMPARs) assembled without the GluR2 subunit. To elucidate the role of these Ca2+-permeable AMPARs, we converted them into Ca2+-impermeable receptors by adenoviral-mediated delivery of the GluR2 gene. This conversion retracted the glial processes ensheathing synapses on Purkinje cell dendritic spines and retarded the removal of synaptically released glutamate. Furthermore, it caused multiple innervation of Purkinje cells by the climbing fibers. Thus, the glial Ca2+-permeable AMPARs are indispensable for proper structural and functional relations between Bergmann glia and glutamatergic synapses.
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              The Cerebellum and Neural Control

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                Author and article information

                Contributors
                Role: ND
                Role: ND
                Journal
                biocell
                Biocell
                Biocell
                Sociedad Latinoamericana de Microscopía Electrónica.; Centro Regional de Investigaciones Científicas y Tecnológicas (Mendoza, Argentina) (Mendoza, Mendoza, Argentina )
                0327-9545
                1667-5746
                August 2009
                : 33
                : 2
                : 71-80
                Affiliations
                [01] Maracaibo orgnameUniversity of Zulia orgdiv1Faculty of Medicine orgdiv2Institute of Biological Investigations Venezuela
                [02] Iowa IA orgnameUniversity of Iowa orgdiv1335 Biology Building orgdiv2Department of Biological Sciences USA
                Article
                S0327-95452009000200001
                fe442801-03c0-4557-bfd1-7b97961f69e6

                This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

                History
                : 12 March 2009
                : 06 April 2009
                : 26 November 2007
                Page count
                Figures: 0, Tables: 0, Equations: 0, References: 61, Pages: 10
                Product

                SciELO Argentina


                Neurons,Confocal microscopy,Glutamate receptors,Glia
                Neurons, Confocal microscopy, Glutamate receptors, Glia

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