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Open Access 01-04-2018 | Original Article

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

Authors: Rafael Luján, Carolina Aguado, Francisco Ciruela, Javier Cózar, David Kleindienst, Luis de la Ossa, Bernhard Bettler, Kevin Wickman, Masahiko Watanabe, Ryuichi Shigemoto, Yugo Fukazawa

Published in: Brain Structure and Function | Issue 3/2018

Abstract

Metabotropic GABA_B receptors mediate slow inhibitory effects presynaptically and postsynaptically through the modulation of different effector signalling pathways. Here, we analysed the distribution of GABA_B receptors using highly sensitive SDS-digested freeze-fracture replica labelling in mouse cerebellar Purkinje cells. Immunoreactivity for GABA_B1 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 immunoparticles increasing 26-fold from somata to dendritic spines. To understand the spatial relationship of GABA_B 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 GABA_B receptors co-assembled with GIRK and Ca_V2.1 channels in the cerebellum. Using double-labelling immunoelectron microscopic techniques, co-clustering between GABA_B1 and GIRK2 was detected in dendritic spines, whereas they were mainly segregated in the dendritic shafts. In contrast, co-clustering of GABA_B1 and Ca_V2.1 was detected in dendritic shafts but not spines. Presynaptically, although no significant co-clustering of GABA_B1 and GIRK2 or Ca_V2.1 channels was detected, inter-cluster distance for GABA_B1 and GIRK2 was significantly smaller in the active zone than in the dendritic shafts, and that for GABA_B1 and Ca_V2.1 was significantly smaller in the active zone than in the dendritic shafts and spines. Thus, GABA_B receptors are associated with GIRK and Ca_V2.1 channels in different subcellular compartments. These data provide a better framework for understanding the different roles played by GABA_B receptors and their effector ion channels in the cerebellar network.

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Title: Differential association of GABAB receptors with their effector ion channels in Purkinje cells
Authors: Rafael Luján
Carolina Aguado
Francisco Ciruela
Javier Cózar
David Kleindienst
Luis de la Ossa
Bernhard Bettler
Kevin Wickman
Masahiko Watanabe
Ryuichi Shigemoto
Yugo Fukazawa
Publication date: 01-04-2018
Publisher: Springer Berlin Heidelberg
Published in: Brain Structure and Function / Issue 3/2018
Print ISSN: 1863-2653
Electronic ISSN: 1863-2661
DOI: https://doi.org/10.1007/s00429-017-1568-y

Keynote webinar | Spotlight on medication adherence

Springer Medicine

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

Abstract

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

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