Co-immunoprecipitation Methods for Detection of G Protein-Coupled Receptors in Brain Tissue
Co-immunoprecipitation Methods for Detection of G Protein-Coupled Receptors in Brain Tissue
复制标题
检测脑组织中 G 蛋白偶联受体的免疫共沉淀方法
DOI：
10.1007/978-1-4939-8985-0_1
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发表时间：
2019
期刊：
影响因子：
--
通讯作者：
Kazunori Namba and Hiroki Kaneko
中科院分区：
文献类型：
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作者：
Nguyen Le Thuy Thi;Takemura Aya J.;Ohniwa Ryosuke L.;Saito Shinji;Morikawa Kazuya;Kazunori Namba and Hiroki Kaneko
一些G蛋白偶联受体(GPCRs)通过与其他类型的GPCRs相互作用来修改自己的信号。在脑组织中，免疫共沉淀是检测GPCR寡聚体最有效的方法之一。脑组织中分子直接相关的证据是可取的，因为体内条件比体外研究更接近自然环境。GPCR异寡聚可能导致新的GPCR靶向药物产生意想不到的效果，使得在体内检测这些组装对药物发现具有重要意义。然而，与体外实验相比，GPCR在脑组织中的表达通常较低，因为启动子被用来人工驱动GPCR的表达；因此，在脑组织中检测到的异构体相互作用较少。因此，含有脑组织免疫共沉淀数据的GPCR异构体寡聚反应的报道很少。在本章中，我们介绍了一种利用啮齿动物脑组织进行GPCRs免疫共沉淀的有效方法。我们修改了一个通用的方案，以获得GPCR在大脑中相互作用的直接证据。
Some G protein-coupled receptors (GPCRs) modify their own signaling by interacting with other GPCR types. In brain tissue, co-immunoprecipitation is one of the most effective method by which to detect GPCR oligomers. Evidence of the direct association of molecules in brain tissue is desirable as in vivo conditions more closely mimic the natural environment than do in vitro studies. GPCR hetero-oligomerization may contribute to the unexpected effects of new GPCR-targeted drugs, making detection of these assemblies in vivo important for drug discovery. However, GPCR expression is generally lower in brain tissue compared with that in in vitro experiments, as promoters are used to artificially drive GPCR expression; consequently, less heteromeric interaction is detectable in brain tissue. Therefore, there are few reports of GPCR heteromeric oligomerization containing co-immunoprecipitation data from brain tissue.In this chapter, we introduce an effective method for co-immunoprecipitation of GPCRs using rodent brain tissue. We modified a general protocol to obtain direct evidence of GPCR interaction in the brain.