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首页> 外文期刊>BMC Neuroscience >Surface clustering of metabotropic glutamate receptor 1 induced by long Homer proteins
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Surface clustering of metabotropic glutamate receptor 1 induced by long Homer proteins

机译：长荷马蛋白诱导的代谢型谷氨酸受体1的表面聚集

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Background Metabotropic glutamate receptors (mGluRs) regulate neuronal excitability and synaptic strength. The group I mGluRs, mGluR1 and 5, are widespread in the brain and localize to post-synaptic sites. The Homer protein family regulates group I mGluR function and distribution. Constitutively expressed 'long' Homer proteins (Homer 1b, 1c, 2 and 3) induce dendritic localization of group I mGluRs and receptor clustering, either internally or on the plasma membrane. Short Homer proteins (Homer 1a, Ania-3) exhibit regulated expression and act as dominant negatives, producing effects on mGluR distribution and function that oppose those of the long Homer proteins. There remains some controversy over whether long Homer proteins induce receptor internalization by inducing retention in the endoplasmic reticulum, or induce mGluR clustering on the plasma membrane. Further, an exhaustive study of the effects of each long Homer isoform on mGluR distribution has not been published. Results The distribution of a GFP-tagged group I mGluR, mGluR1-GFP, was examined in the absence of Homer proteins and in the presence of several Homer isoforms expressed in sympathetic neurons from the rat superior cervical ganglion (SCG) using total internal reflection fluorescence (TIRF-M) and confocal microscopy. Quantitative analysis of mGluR1-GFP fluorescence using TIRF-M revealed that expression of each long Homer isoform tested (Homer 1b, 1c, 2b and 3) induced a significant degree of surface clustering. Using confocal imaging, Homer-induced mGluR clusters were observed intra-cellularly as well as on the plasma membrane. Further, in approximately 40% of neurons co-expressing mGluR1-GFP and Homer 1b, intracellular inclusions were observed, but plasma membrane clusters were also documented in some Homer 1b coexpressing cells. Conclusion All long Homer proteins examined (Homer 1b, 1c, 2b and 3) induced a significant degree of mGluR1-GFP clustering on the plasma membrane compared to cells expressing mGluR1-GFP alone. Clusters induced by long Homers appeared on the plasma membrane and intracellularly, suggesting that clusters form prior to plasma membrane insertion and/or persist after internalization. Finally, while Homer 1b induced surface clustering of mGluR1 in some cells, under some conditions intracellular retention may occur.
机译：背景代谢型谷氨酸受体（mGluRs）调节神经元兴奋性和突触强度。第一组的mGluR，mGluR1和5在大脑中广泛分布，并位于突触后部位。荷马蛋白家族调节I组mGluR的功能和分布。组成型表达的“长”荷马蛋白（荷马1b，1c，2和3）在内部或在质膜上诱导I组mGluRs的树突状定位和受体簇集。短的荷马蛋白（荷马1a，Ania-3）表现出调节的表达，并作为显性负片，对mGluR分布和功能产生影响，而长荷马蛋白则与之相反。关于长的荷马蛋白是通过诱导内质网中的滞留来诱导受体内在化，还是在质膜上诱导mGluR聚集，仍然存在争议。此外，尚未发表对每种长的荷马异构体对mGluR分布的影响的详尽研究。结果使用全内反射荧光在没有荷马蛋白的情况下和在大鼠上颈神经节（SCG）的交感神经元中表达的几种荷马亚型的存在下，检查了带有GFP标签的I组mGluR，mGluR1-GFP的分布。 （TIRF-M）和共聚焦显微镜检查。使用TIRF-M对mGluR1-GFP荧光进行的定量分析显示，每种长的荷马同种型（荷马1b，1c，2b和3）的表达都诱导了很大程度的表面聚集。使用共聚焦成像，在细胞内以及质膜上观察到荷马诱导的mGluR簇。此外，在约40％的共表达mGluR1-GFP和Homer 1b的神经元中，观察到细胞内包涵体，但在某些Homer 1b共表达细胞中也记录到质膜簇。结论与仅表达mGluR1-GFP的细胞相比，所检查的所有较长的Homer蛋白（荷马1b，1c，2b和3）均在质膜上诱导了显着程度的mGluR1-GFP聚集。由长霍默斯诱导的簇出现在质膜上和细胞内，表明该簇在质膜插入之前形成和/或在内化后持续存在。最后，尽管荷马1b在某些细胞中诱导了mGluR1的表面聚集，但在某些条件下可能会发生细胞内滞留。

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