Cellular distribution of the rat D-4 dopamine receptor protein in the CNS using anti-receptor antisera
Cellular distribution of the rat D-4 dopamine receptor protein in the CNS using anti-receptor antisera
复制标题
用抗受体抗血清检测大鼠多巴胺受体蛋白在中枢神经系统中的细胞分布
DOI：
10.1016/s0006-8993(96)01422-9
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发表时间：
1997-03-28
期刊：
影响因子：
2.9
通讯作者：
Sibley, DR
中科院分区：
文献类型：
--
作者：
Ariano, MA;Wang, J;Sibley, DR
针对大鼠D-4多巴胺（DA）受体中的独特肽段产生多克隆抗血清。使用固相ELISA监测滴度，一旦确定，使用稳定转染大鼠D-4 DA受体全长cDNA的中国卵巢（CHO）细胞评估特异性。与抗D-4 DA受体抗血清孵育产生的免疫荧光染色对D-4 DA受体转染的CHO细胞具有选择性，并在其细胞膜和细胞质中表达。在未转染的K1 CHO细胞和D-2或D-3 DA受体转染的CHO细胞中，可见D-4 DA受体蛋白的减弱染色。检查了D-4、DA受体亚型的区域和细胞CNS分布模式，并说明了额叶（FCx）和顶叶皮质内的显著蛋白水平。在丘脑、苍白球、海马、小脑蚓部中出现少量受体蛋白染色，在纹状体（CPu）中检测到非常低的表达。使用同步荧光分析，D-4 DA受体蛋白染色与这些相同脑区中其mRNA转录本的细胞表达相关。D-4 DA受体转录本和编码蛋白在FCx和CPu的神经元中的染色模式的同源性一致性表明，在这些确定的基底神经节通路中受体表达存在差异。(C)1997年Elsevier Science B.V.
A polyclonal antiserum was generated against a unique peptide fragment in the rat D-4 dopamine (DA) receptor. The titer was monitored using solid-phase ELISA and once it was established, specificity was assessed using Chinese Hamster Ovary (CHO) cells, stably transfected with the full-length cDNA for the rat D-4 DA receptor. Immunofluorescent staining produced by incubation with the anti-D-4 DA receptor antiserum was selective for D-4 DA receptor-transfected CHO cells, and was expressed at their cell membranes and cytoplasm. Attenuated staining for D-4 DA receptor protein was visible in untransfected, K1 CHO cells, and in D-2 or D-3 DA receptor-transfected CHO cells. The regional and cellular CNS distribution patterns for the D-4, DA receptor subtype were examined, and illustrated significant protein levels within the frontal (FCx) and parietal cortices. Lesser amounts of receptor protein staining occurred in the thalamus, globus pallidus, hippocampus, cerebellar vermis, and very low expression was detected in the striatum (CPu). D-4 DA receptor protein staining was correlated with the cellular expression of its mRNA transcripts in these same brain regions using concurrent fluorescent analyses. The homologous coincidence in staining patterns for the D-4 DA receptor transcripts and encoded proteins in identified neurons of the FCx and CPu showed variations in receptor expression in these identified basal ganglia pathways. (C) 1997 Elsevier Science B.V.