Expression of alpha subunit genes of nicotinic acetylcholine receptors in human lymphocytes
Expression of alpha subunit genes of nicotinic acetylcholine receptors in human lymphocytes
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DOI：
10.1016/0304-3940(96)12908-6
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发表时间：
1996-08-23
影响因子：
2.5
通讯作者：
Schroder, H
中科院分区：
文献类型：
--
作者：
Hiemke, C;Stolp, M;Schroder, H
利用免疫组织化学和原位杂交技术，我们研究了烟碱乙酰胆碱受体(nAChRs) α亚基是否在人淋巴细胞中表达。采用差速低速梯度离心从10名健康志愿者肝素化静脉血中分离细胞。受体位点使用单克隆抗体WF6可视化，该抗体特异性识别包括人类在内的几种物种的α -亚型。为了可视化转录本，使用地高辛标记的α 4-和α 3亚基的cRNA探针。免疫染色显示WF6与分离的人淋巴样细胞特异性结合。该抗体与大多数细胞结合，并优先集中在核周/表面区域。免疫反应性与应用一种识别CD4表面蛋白的抗体进行比较后观察到的相似。原位杂交结果显示，nAChRs α 4亚基基因在所有先显子的淋巴细胞中均有表达。在10个个体中，有8个个体发现α 3亚基的强度低于α 4转录本。带有相应意义rna的对照孵育结果为阴性。由此可见，人类淋巴细胞能够表达nachr的α亚基基因。
Using immunohistochemistry and in situ hybridisation, we have studied whether alpha-subunits of nicotinic acetylcholine receptors (nAChRs) are expressed in human lymphocytes. Cells were isolated by differential low speed gradient centrifugation from heparinised venous blood of 10 healthy volunteers. Receptor sites were visualised using the monoclonal antibody WF6 which specifically recognises alpha-isoforms from several species including man. For visualisation of transcripts, digoxigenin-labelled cRNA probes for alpha 4- and alpha 3-subunits were used. Immunostaining revealed specific binding of WF6 to isolated human lymphoid cells. The antibody was bound to most cells and concentrated preferentially in the perinuclear/surface region. The immunoreactivity resembled that observed after application of an antibody recognising CD4 surface proteins which was conducted for comparison. In situ-hybridisation revealed that the alpha 4-subunit genes of nAChRs was expressed in lymphocytes of all probands. The alpha 3-subunit was found, with lower intensity than alpha 4-transcripts, in eight of the 10 individuals. Control incubations with corresponding sense cRNAs were negative. It is concluded that human lymphocytes are able to express alpha-subunit genes of nAChRs.