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代谢型谷氨酸受体1的mGluR1b剪接变体在大鼠小脑皮质平行纤维-浦肯野细胞突触处的免疫定位。

Immunolocalization of the mGluR1b splice variant of the metabotropic glutamate receptor 1 at parallel fiber-Purkinje cell synapses in the rat cerebellar cortex.

作者信息

Mateos J M, Benítez R, Elezgarai I, Azkue J J, Lázaro E, Osorio A, Bilbao A, Doñate F, Sarría R, Conquet F, Ferraguti F, Kuhn R, Knöpfel T, Grandes P

机构信息

Department of Neurosciences, Faculty of Medicine and Dentistry, Basque Country University, Bilbao, Spain.

出版信息

J Neurochem. 2000 Mar;74(3):1301-9. doi: 10.1046/j.1471-4159.2000.741301.x.

Abstract

Several metabotropic glutamate receptor (mGluR) subtypes have been identified in the cerebellar cortex that are targeted to different compartments in cerebellar cells. In this study, preembedding immunocytochemical methods for electron microscopy were used to investigate the subcellular distribution of the mGluR1b splice variant in the rat cerebellar cortex. Dendritic spines of Purkinje cells receiving parallel fiber synaptic terminals were immunoreactive for mGluR1b. With a preembedding immunogold method, approximately 25% of the mGluR1b immunolabeling was observed perisynaptically within 60 nm from the edge of the postsynaptic densities. Values of extrasynaptic gold particles beyond the first 60 nm were maintained at between 10 and 18% along the whole intracellular surface of the dendritic spine membranes of Purkinje cells. For comparison, the distribution of mGluR1a was studied. A predominant (approximately 37%) perisynaptic localization of mGluR1a was seen in dendritic spines of Purkinje cells, dropping the extrasynaptic labeling to 15% in the 60-120-nm bin from the edge of the postsynaptic specialization. Our results reveal that mGluR1b and mGluR1a are localized to the same subcellular compartments in Purkinje cells but that the densities of the perisynaptic and extrasynaptic pools were different for both isoforms. The compartmentalization of mGluR1b and mGluR1a might serve distinct requirements in cerebellar neurotransmission.

摘要

在小脑皮质中已鉴定出几种代谢型谷氨酸受体(mGluR)亚型,它们定位于小脑细胞的不同区域。在本研究中,采用用于电子显微镜的包埋前免疫细胞化学方法,研究大鼠小脑皮质中mGluR1b剪接变体的亚细胞分布。接受平行纤维突触终末的浦肯野细胞的树突棘对mGluR1b具有免疫反应性。采用包埋前免疫金法,在距突触后致密物边缘60nm内的突触周围观察到约25%的mGluR1b免疫标记。在浦肯野细胞树突棘膜的整个细胞内表面,超出最初60nm的突触外金颗粒值维持在10%至18%之间。为作比较,研究了mGluR1a的分布。在浦肯野细胞的树突棘中观察到mGluR1a主要(约37%)定位于突触周围,使突触后特化边缘60-120nm区间内的突触外标记降至15%。我们的结果表明,mGluR1b和mGluR1a定位于浦肯野细胞的相同亚细胞区域,但两种亚型的突触周围池和突触外池的密度不同。mGluR1b和mGluR1a的区室化可能在小脑神经传递中满足不同的需求。

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