Differential association of GABA(B) receptors with their effector ion channels in Purkinje cells.
Differential association of GABA(B) receptors with their effector ion channels in Purkinje cells.
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DOI：
10.1007/s00429-017-1568-y
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发表时间：
2018-04
影响因子：
3.1
通讯作者：
Fukazawa Y
中科院分区：
文献类型：
--
作者：
Luján R;Aguado C;Ciruela F;Cózar J;Kleindienst D;de la Ossa L;Bettler B;Wickman K;Watanabe M;Shigemoto R;Fukazawa Y
代谢型GABAB受体通过调节不同的效应器信号传导途径在突触前和突触后介导缓慢的抑制作用。在这里，我们分析了GABAB受体的分布，使用高度敏感的SDS消化冷冻骨折复制品标记在小鼠小脑浦肯野细胞。GABAB 1的免疫反应性，观察突触前，更丰富，突触后车厢，显示分散和集群的分布模式。免疫颗粒的定量分析揭示了体细胞-树突梯度，从体细胞到树突棘免疫颗粒的密度增加了26倍。为了了解GABAB受体与两个关键效应离子通道（G蛋白门控内向整流K+（GIRK/Kir 3）通道和电压依赖性Ca 2+通道）的空间关系，进行了生化和免疫组织化学方法。免疫共沉淀分析表明GABAB受体与小脑中的GIRK和CaV2.1通道共组装。使用双标记免疫电镜技术，GABAB 1和GIRK 2之间的共聚类检测到树突棘，而他们主要是在树突状细胞的轴分离。与此相反，GABAB 1和CaV2.1的共聚集检测到树突状干，但不是刺。突触前，虽然没有显着的GABAB 1和GIRK 2或CaV2.1通道的共聚类检测，GABAB 1和GIRK 2的簇间距离是显着小于在活动区比在树突状干，和GABAB 1和CaV2.1是显着小于在活动区比在树突状干和棘。因此，GABAB受体与不同亚细胞区室中的GIRK和CaV2.1通道相关。这些数据为了解GABAB受体及其效应离子通道在小脑网络中发挥的不同作用提供了更好的框架。
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 immunoparticles 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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影响因子：
3.7
作者：
Degro CE;Kulik A;Booker SA;Vida I
通讯作者：
Vida I
DOI：
10.1523/jneurosci.2921-12.2013
发表时间：
2013-02-20
期刊：
The Journal of neuroscience : the official journal of the Society for Neuroscience
影响因子：
--
作者：
Indriati DW;Kamasawa N;Matsui K;Meredith AL;Watanabe M;Shigemoto R
通讯作者：
Shigemoto R
影响因子：
2.5
作者：
Fritschy, JM;Sidler, C;Benke, D
通讯作者：
Benke, D
影响因子：
3.5
作者：
Clancy, SM;Fowler, CE;Slesinger, PA
通讯作者：
Slesinger, PA
影响因子：
3.3
作者：
HUSTON, E;CULLEN, GP;DOLPHIN, AC
通讯作者：
DOLPHIN, AC