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首页> 外文期刊>The European Journal of Neuroscience >Distinct subsynaptic localization of type 1 metabotropic glutamate receptors at glutamatergic and GABAergic synapses in the rodent cerebellar cortex
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Distinct subsynaptic localization of type 1 metabotropic glutamate receptors at glutamatergic and GABAergic synapses in the rodent cerebellar cortex

机译：啮齿类小脑皮层中谷氨酸能和GABA能突触的1型代谢型谷氨酸受体的不同突触亚定位

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摘要

Type 1 metabotropic glutamate (mGlu1) receptors play a pivotal role in different forms of synaptic plasticity in the cerebellar cortex, e.g. long-term depression at glutamatergic synapses and rebound potentiation at GABAergic synapses. These various forms of plasticity might depend on the subsynaptic arrangement of the receptor in Purkinje cells that can be regulated by protein-protein interactions. This study investigated, by means of the freeze-fracture replica immunogold labelling method, the subcellular localization of mGlu1 receptors in the rodent cerebellum and whether Homer proteins regulate their subsynaptic distribution. We observed a widespread extrasynaptic localization of mGlu1 receptors and confirmed their peri-synaptic enrichment at glutamatergic synapses. Conversely, we detected mGlu1 receptors within the main body of GABAergic synapses onto Purkinje cell dendrites. Although Homer proteins are known to interact with the mGlu1 receptor C-terminus, we could not detect Homer3, the most abundant Homer protein in the cerebellar cortex, at GABAergic synapses by pre-embedding and post-embedding immunoelectron microscopy. We then hypothesized a critical role for Homer proteins in the peri-junctional localization of mGlu1 receptors at glutamatergic synapses. To disrupt Homer-associated protein complexes, mice were tail-vein injected with the membrane-permeable dominant-negative TAT-Homer1a. Freeze-fracture replica immunogold labelling analysis showed no significant alteration in the mGlu1 receptor distribution pattern at parallel fibre-Purkinje cell synapses, suggesting that other scaffolding proteins are involved in the peri-synaptic confinement. The identification of interactors that regulate the subsynaptic localization of the mGlu1 receptor at neurochemically distinct synapses may offer new insight into its trafficking and intracellular signalling.
机译：1型代谢型谷氨酸（mGlu1）受体在小脑皮层的不同形式的突触可塑性中起关键作用，例如谷氨酸能突触的长期抑郁和GABA能突触的反弹增强。这些各种形式的可塑性可能取决于浦肯野细胞中受体的突触下排列，该排列可以通过蛋白质-蛋白质相互作用来调节。该研究通过冷冻断裂复制免疫金标记方法研究了啮齿类小脑中mGlu1受体的亚细胞定位以及Homer蛋白是否调节其突触下分布。我们观察到一个广泛的mGlu1受体突触外定位，并确认他们在谷氨酸能突触周围突触丰富。相反地​​，我们在Purkinje细胞树突上检测到了GABA能突触的主体内的mGlu1受体。尽管已知荷马蛋白与mGlu1受体C末端相互作用，但我们无法通过嵌入前和嵌入后免疫电子显微镜在GABA能突触中检测到小脑皮质中最丰富的荷马蛋白Homer3。然后，我们假设荷马蛋白在谷氨酸能突触处的mGlu1受体的周围连接局部化中起关键作用。为了破坏荷马相关蛋白复合物，向小鼠尾静脉注射膜可渗透的显性负性TAT-Homer1a。冷冻断裂复制品免疫金标记分析显示平行纤维-Purkinje细胞突触处的mGlu1受体分布模式没有显着改变，表明其他支架蛋白参与突触周围的封闭。调节mGlu1受体在神经化学上不同突触的突触下的定位的相互作用的鉴定可能提供新的见解其贩运和细胞内信号。

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