Evidence against dopamine D1/D2 receptor heteromers.
Evidence against dopamine D1/D2 receptor heteromers.
复制标题
DOI：
10.1038/mp.2014.166
复制
发表时间：
2015-11
影响因子：
11
通讯作者：
Javitch, J. A.
中科院分区：
文献类型：
--
作者：
Frederick, A. L.;Yano, H.;Trifilieff, P.;Vishwasrao, H. D.;Biezonski, D.;Meszaros, J.;Urizar, E.;Sibley, D. R.;Kellendonk, C.;Sonntag, K. C.;Graham, D. L.;Colbran, R. J.;Stanwood, G. D.;Javitch, J. A.
G蛋白偶联受体的异位低聚物已成为研究的热点，因为其潜在的信号和药理特性不同于组份受体，这使得它们在开发更具选择性的药物治疗方面具有极大的吸引力。特别是，多巴胺D1和D2受体可以形成与GαQ蛋白偶联的异源低聚物，SKF83959被认为是一种有偏向的激动剂，选择性地与这些受体复合体结合来激活GαQ，因此磷脂酶C.D1/D2异构体被认为与抑郁症和精神分裂症的病理生理和治疗有关。我们使用了体外生物发光共振能量转移(BRET)、体外分析脑片中受体的定位和接近程度以及小鼠的行为分析来表征来自这些假定的二聚体/低聚物的信号。我们无法用多种生物传感器检测到与D_1或D_2受体的同或异构体偶联的G-α-Q或G-α-11蛋白。在D1R基因敲除小鼠中，SKF83959诱导的运动和梳理行为被消除，证实了D1样受体激活的关键作用。相比之下，SKF83959诱导的运动反应在D2受体和GαQ基因敲除小鼠中是完整的，在表达突变的Ala286-CaMKIIα的敲入小鼠中也是如此，该突变不能自动磷酸化而变得活跃。此外，我们发现在伏核的外壳中，即使在D1和D2受体启动子都活跃的神经元中，受体蛋白也是分离的，不形成复合体。这些数据与SKF83959通过GαQ或通过D1-D2异构体的信号不兼容，并挑战了我们在研究中使用的成年动物中这种信号复合体的存在。
Hetero-oligomers of G-protein-coupled receptors have become the subject of intense investigation because their purported potential to manifest signaling and pharmacological properties that differ from the component receptors makes them highly attractive for the development of more selective pharmacological treatments. In particular, dopamine D1 and D2 receptors have been proposed to form hetero-oligomers that couple to Gαq proteins, and SKF83959 has been proposed to act as a biased agonist that selectively engages these receptor complexes to activate Gαq and thus phospholipase C. D1/D2 heteromers have been proposed as relevant to the pathophysiology and treatment of depression and schizophrenia. We used in vitro bioluminescence resonance energy transfer (BRET), ex vivo analyses of receptor localization and proximity in brain slices, and behavioral assays in mice to characterize signaling from these putative dimers/oligomers. We were unable to detect Gαq or Gα11 protein coupling to homomers or heteromers of D1 or D2 receptors using a variety of biosensors. SKF83959-induced locomotor and grooming behaviors were eliminated in D1 receptor knockout mice, verifying a key role for D1-like receptor activation. In contrast, SKF83959-induced motor responses were intact in D2 receptor and Gαq knockout mice, as well as in knock-in mice expressing a mutant Ala286-CaMKIIα, that cannot autophosphorylate to become active. Moreover, we found that in the shell of the nucleus accumbens, even in neurons in which D1 and D2 receptor promoters are both active, the receptor proteins are segregated and do not form complexes. These data are not compatible with SKF83959 signaling through Gαq or through a D1–D2 heteromer and challenge the existence of such a signaling complex in the adult animals that we used for our studies.
登录
查看更多内容
通过逆行标记鉴定纹状体投射神经元类型上 D1 和 D2 多巴胺受体的大鼠核周差异定位
影响因子：
2.8
作者：
Deng, Yun-Ping;Lei, Wan-Long;Reiner, Anton
通讯作者：
Reiner, Anton
影响因子：
3
作者：
Frederick AL;Saborido TP;Stanwood GD
通讯作者：
Stanwood GD
影响因子：
5
作者：
ARNT, J;HYTTEL, J;SANCHEZ, C
通讯作者：
SANCHEZ, C
影响因子：
3.6
作者：
Chun, Lani S.;Free, R. Benjamin;Sibley, David R.
通讯作者：
Sibley, David R.
影响因子：
5
作者：
Andringa, G;Drukarch, B;Stoof, JC
通讯作者：
Stoof, JC