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    Differential association of GABAB receptors with their effector ion channels in Purkinje cells

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    Versión post-print (5.135Mb)
    Fecha
    2017-11-25
    Autor
    Luján Miras, Rafael
    Aguado, Carolina
    Ciruela, Francisco
    Cozar del Olmo, Javier
    Kleindienst, David
    Ossa, Luis de la
    Bettler, Bernhard
    Wickman, Kevin
    Watanabe, Masahiko
    Shigemoto, Ryuichi
    Fukazawa, Yugo
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    Resumen
    Metabotropic GABAB receptors mediate slow inhibitory effects presynaptically and postsynaptically through the modulation of different effector signalling pathways. Here, we analysed the distribution of GABAB receptors using highly sensitive SDS-digested freeze-fracture replica labelling in mouse cerebellar Purkinje cells. Immunoreactivity for GABAB1 was observed on presynaptic and, more abundantly, on postsynaptic compartments, showing both scattered and clustered distribution patterns. Quantitative analysis of immunoparticles revealed a somato-dendritic gradient, with the density of mmunoparticles increasing 26-fold from somata to dendritic spines. To understand the spatial relationship of GABAB receptors with two key effector ion channels, the G protein-gated inwardly rectifying K? (GIRK/Kir3) channel and the voltage-dependent Ca2? channel, biochemical and immunohistochemical approaches were performed. Co-immunoprecipitation analysis demonstrated that GABAB receptors co-assembled with GIRK and CaV2.1 channels in the cerebellum. Using double-labelling immunoelectron microscopic techniques, co-clustering between GABAB1 and GIRK2 was detected in dendritic spines, whereas they were mainly segregated in the dendritic shafts. In contrast, co-clustering of GABAB1 and CaV2.1 was detected in dendritic shafts but not spines. Presynaptically, although no significant co-clustering of GABAB1 and GIRK2 or CaV2.1 channels was detected, inter-cluster distance for GABAB1 and GIRK2 was significantly smaller in the active zone than in the dendritic shafts, and that for GABAB1 and CaV2.1 was significantly smaller in the active zone than in the dendritic shafts and spines. Thus, GABAB receptors are associated with GIRK and CaV2.1 channels in different subcellular compartments. These data provide a better framework for understanding the different roles played by GABAB receptors and their effector ion channels in the cerebellar network.
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    http://hdl.handle.net/10578/20973
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    • Área de Histología

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    © Universidad de Castilla-La Mancha
    Rectorado
    C/ Altagracia, 50 13071
    Ciudad Real Tfno. 926 29 53 00
    Fax: 926 29 53 01

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