Department of Neuroscience, Columbia University, New York, NY, USA. pt2194@columbia
Biotechniques. 2011 Aug;51(2):111-8. doi: 10.2144/000113719.
The existence of G protein-coupled receptor (GPCR) dimers and/or oligomers has been demonstrated in heterologous systems using a variety of biochemical and biophysical assays. While these interactions are the subject of intense research because of their potential role in modulating signaling and altering pharmacology, evidence for the existence of receptor interactions in vivo is still elusive because of a lack of appropriate methods to detect them. Here, we adapted and optimized a proximity ligation assay (PLA) for the detection in brain slices of molecular proximity of two antigens located on either the same or two different GPCRs. Using this approach, we were able to confirm the existence of dopamine D2 and adenosine A2A receptor complexes in the striatum of mice ex vivo.
G 蛋白偶联受体 (GPCR) 二聚体和/或寡聚体的存在已在使用各种生化和生物物理测定法的异源系统中得到证实。虽然这些相互作用是研究的热点，因为它们可能在调节信号转导和改变药理学方面发挥作用，但由于缺乏适当的方法来检测它们，体内受体相互作用的证据仍然难以捉摸。在这里，我们对一种接近性连接测定法 (PLA) 进行了改编和优化，以检测位于同一或两个不同 GPCR 上的两种抗原在脑切片中的分子接近程度。使用这种方法，我们能够在体外确认小鼠纹状体中多巴胺 D2 和腺苷 A2A 受体复合物的存在。
