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实时荧光定量PCR检测人外周血淋巴细胞多巴胺基因受体(DRD1-DRD5)的表达

The Detection of Dopamine Gene Receptors (DRD1-DRD5) Expression on Human Peripheral Blood Lymphocytes by Real Time PCR.

作者信息

Ostadali Mohammad Reza, Ahangari Ghasem, Eslami Mohammad Bagher, Razavi Alireza, Zarrindast Mohammad Reza, Ahmadkhaniha Hamid Reza, Boulhari Jafar

机构信息

Department of Immunology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.

出版信息

Iran J Allergy Asthma Immunol. 2004 Dec;3(4):169-74.

Abstract

There is interrelationship between the immune and nervous systems that is accomplished by the molecular mediators. Dopamine is one of the most important neurotransmitters. Five different dopamine receptor genes (DRD1, DRD2, DRD3, DRD4, and DRD5) have been recognized and cloned. The expression of the dopamine receptors is well characterized in the brain but little work has been done to examine their expression in other organ tissues. In certain diseases of the immune and nervous systems, alterations in dopamine receptors gene expression in different cells have been reported. This suggests that dopamine and its receptors have important role in pathophysiology of above-mentioned diseases.In the present study, using Real Time Polymerase Chain Reaction (PCR) technique, we investigated dopamine receptors genes expression in PBMC of normal individuals. The PBMC was separated from normal whole blood by Ficoll-hypaque; the total cellular RNA was then extracted and the cDNA was synthesized. This process followed by real time-PCR using primer pairs specific for five dopamine receptors mRNAs and beta-actin as internal control. The results showed the presence of all types of dopamine receptors in lymphocytes of normal individuals. The specificities of the obtained PCR products for the respective dopamine receptors fragments were confirmed by sequenced analysis capillary system. In conclusion, the present study has shown that human lymphocytes express five dopamine receptors DR1-DR5. However, the conclusive evidence on the possible function of these receptors in lymphocytes remains unknown. Because lymphocytes express all of the five neuronal dopamine receptors, it is quite reasonable to consider them as a model of dopaminergic neuron.

摘要

免疫系统和神经系统之间存在着由分子介质介导的相互关系。多巴胺是最重要的神经递质之一。已识别并克隆出五种不同的多巴胺受体基因(DRD1、DRD2、DRD3、DRD4和DRD5)。多巴胺受体在大脑中的表达已得到充分表征,但在其他器官组织中研究其表达的工作却很少。在某些免疫和神经系统疾病中,已报道不同细胞中多巴胺受体基因表达的改变。这表明多巴胺及其受体在上述疾病的病理生理学中具有重要作用。在本研究中,我们使用实时聚合酶链反应(PCR)技术,研究了正常个体外周血单个核细胞(PBMC)中多巴胺受体基因的表达。通过Ficoll-淋巴细胞分离液从正常全血中分离出PBMC;然后提取总细胞RNA并合成cDNA。此过程之后,使用针对五种多巴胺受体mRNA的引物对和β-肌动蛋白作为内对照进行实时PCR。结果显示正常个体的淋巴细胞中存在所有类型的多巴胺受体。通过测序分析毛细管系统确认了所获得的PCR产物对各个多巴胺受体片段的特异性。总之,本研究表明人类淋巴细胞表达五种多巴胺受体DR1-DR5。然而,这些受体在淋巴细胞中可能功能的确凿证据仍然未知。由于淋巴细胞表达所有五种神经元多巴胺受体,将它们视为多巴胺能神经元的模型是相当合理的。

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