LOCALIZATION OF D(1) AND D(2) DOPAMINE-RECEPTORS IN BRAIN WITH SUBTYPE-SPECIFIC ANTIBODIES
LOCALIZATION OF D(1) AND D(2) DOPAMINE-RECEPTORS IN BRAIN WITH SUBTYPE-SPECIFIC ANTIBODIES
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DOI：
10.1073/pnas.90.19.8861
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发表时间：
1993-10-01
影响因子：
11.1
通讯作者：
CILIAX, BJ
中科院分区：
文献类型：
--
作者：
LEVEY, AI;HERSCH, SM;CILIAX, BJ
五个或更多的多巴胺受体基因在大脑中表达。然而，编码的D1-D5受体的药理学相似性阻碍了对其亚型的定位和功能的研究。为了更好地了解单个受体的作用，人们提出了针对重组D1和D2蛋白的抗体，并在Western印迹和免疫沉淀研究中证明抗体能与受体亚型特异结合。每一种抗体都选择性地与各自的受体蛋白发生反应，这些受体蛋白既在cDNA转基因细胞中表达，也在脑中表达。免疫细胞化学显示，D1和D2在大鼠、猴和人脑中的区域分布相似，其中纹状体、嗅球和黑质的染色最强。然而，在每个地区，每个亚型的准确分布是不同的，而且往往是互补的。D1和D2在纹状体斑块和基质隔区、嗅球的选择性层、黑质致密质或网状结构中均有不同程度的丰富。电子显微镜显示，d1和d2也具有高度选择性的亚细胞分布。在大鼠新纹状体，D_1和D_2免疫反应主要定位于大鼠新纹状体内树突和棘头亚群的突触后部位。突触前D_1和D_2受体也被观察到，表明这两个亚型都可能调节神经递质的释放。在黑质的轴突终末也有D1的表达。这些结果为理解哺乳动物大脑中离散神经元回路中遗传决定的D1和D2受体的突触前和突触后功能提供了形态基础。
Five or more dopamine receptor genes are expressed in brain. However, the pharmacological similarities of the encoded D1-D5 receptors have hindered studies of the localization and functions of the subtypes. To better understand the roles of the individual receptors, antibodies were raised against recombinant D1 and D2 proteins and were shown to bind to the receptor subtypes specifically in Western blot and immunoprecipitation studies. Each antibody reacted selectively with the respective receptor protein expressed both in cells transfected with the cDNAs and in brain. By immunocytochemistry, D1 and D2 had similar regional distributions in rat, monkey, and human brain, with the most intense staining in striatum, olfactory bulb, and substantia nigra. Within each region, however, the precise distributions of each subtype were distinct and often complementary. D1 and D2 were differentially enriched in striatal patch and matrix compartments, in selective layers of the olfactory bulb, and in either substantia nigra pan compacta or reticulata. Electron microscopy demonstrated that D1 and D2 also had highly selective subcellular distributions. In the rat neostriatum, the majority of D1 and D2 immunoreactivity was localized in postsynaptic sites in subsets of spiny dendrites and spine heads in rat neostriatum. Presynaptic D1 and D2 receptors were also observed, indicating both subtypes may regulate neurotransmitter release. D1 was also Present in axon terminals in the substantia nigra. These results provide a morphological substrate for understanding the pre- and postsynaptic functions of the genetically defined D1 and D2 receptors in discrete neuronal circuits in mammalian brain.