Choi W S, Machida C A, Ronnekleiv O K
Division of Neuroscience, Oregon Regional Primate Research Center, Beaverton 97006, USA.
Brain Res Mol Brain Res. 1995 Jul;31(1-2):86-94. doi: 10.1016/0169-328x(95)00038-t.
The distribution of the mRNAs encoding the dopamine D1, D2 and D5 receptors was determined in brain tissues obtained from intact female rhesus monkeys, using a ribonuclease protection assay. Tissue blocks from the frontal cortex, striatum, thalamus, hippocampus and substantia nigra were dissected and total RNA was extracted. Dopamine D2 and D5 receptor DNA fragments were generated from rhesus monkey genomic DNA using polymerase chain reaction. To generate dopamine receptor subtype-specific cRNA probes, DNA fragments corresponding to the carboxy terminus of the rhesus monkey D1 and D2 receptor genes and to the putative transmembrane domain regions (IV-VI) of the D5 receptor gene, were subcloned into the pGEM3Z/4Z vectors. Expression of D1 receptor mRNA exhibited significant regional differences: striatum > > > cerebral cortex > or = hippocampus > or = lateral thalamus. D1 receptor mRNA was found in low quantities in the medial thalamus, but was not consistently expressed in the substantia nigra area. In contrast, D2 receptor mRNA was detected in all regions that were studied: striatum > > > substantia nigra > > hippocampus > or = cerebral cortex > or = medial thalamus > or = lateral thalamus. D5 receptor mRNA was also expressed in all regions, with highest levels in the cerebral cortex, striatum and lateral thalamus, and moderate levels in the substantia nigra, medial thalamus and the hippocampus. The D5 receptor mRNA appears to be widely distributed in the monkey brain. Most interesting is the expression of D5 receptor mRNA in tissues of the substantia nigra area.
采用核糖核酸酶保护分析法，测定了从完整雌性恒河猴获取的脑组织中编码多巴胺D1、D2和D5受体的mRNA的分布情况。解剖额叶皮质、纹状体、丘脑、海马体和黑质的组织块并提取总RNA。利用聚合酶链反应从恒河猴基因组DNA中生成多巴胺D2和D5受体DNA片段。为了生成多巴胺受体亚型特异性的cRNA探针，将与恒河猴D1和D2受体基因羧基末端以及D5受体基因假定跨膜结构域区域（IV-VI）对应的DNA片段亚克隆到pGEM3Z/4Z载体中。D1受体mRNA的表达呈现出显著的区域差异：纹状体＞＞＞大脑皮质≥海马体≥外侧丘脑。在内侧丘脑中发现少量D1受体mRNA，但在黑质区域未持续表达。相比之下，在所有研究区域均检测到D2受体mRNA：纹状体＞＞＞黑质＞＞海马体≥大脑皮质≥内侧丘脑≥外侧丘脑。D5受体mRNA在所有区域也均有表达，在大脑皮质、纹状体和外侧丘脑中水平最高，在黑质、内侧丘脑和海马体中水平中等。D5受体mRNA似乎在猴脑中广泛分布。最有意思的是D5受体mRNA在黑质区域组织中的表达。
