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首页> 外文期刊>Life sciences >Intracellular localization of nicotinic acetylcholine receptors in human cell lines.
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Intracellular localization of nicotinic acetylcholine receptors in human cell lines.

机译：人细胞系中烟碱乙酰胆碱受体的细胞内定位。

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Previously we demonstrated that mouse liver mitochondria express functional α7 nicotinic acetylcholine receptors (nAChRs). The aim of this study was to investigate whether the nAChRs are found in mitochondria of non-neuronal human cell lines.Three cell lines: U373 (astrocytes), U937 (monocytes) and Daudi (B lymphocytes) were examined by flow cytometry, Cell ELISA and fluorescent confocal microscopy using the antibodies against extracellular epitopes of α3, α4, α7, α9, β2 and β4 nAChR subunits.It is shown that the studied cells expressed different sets of nAChR subunits on the plasma membrane suggesting the presence of α7 nAChRs on all cells, of α3β4 nAChRs on U373 cells and of α4β2/α4β4 nAChRs on U937 cells. In addition to nAChRs exposed on the surface, all cells contained a considerable intracellular pool of α3- and α7-containing nAChRs. The binding of α3-, α7- and β4-specific antibodies partially overlapped with that of mitochondrial outer membrane translocase-specific antibody. Binding of α7-specific antibody also overlapped with that of MitoTracker Green, which binds to active mitochondria.The data obtained suggest that a part of intracellular α3β4 and α7 nAChRs in U373, U937 and Daudi cells is located on mitochondria. This finding complements our previous observation of α7 nAChRs in mouse liver mitochondria and reveals new intracellular targets for cholinergic regulation.
机译：以前我们证明小鼠肝线粒体表达官能α7烟碱乙酰胆碱受体（NACHRS）。本研究的目的是调查NACHRS是否发现在非神经元人细胞系的线粒体中。通过流式细胞术，细胞ELISA检查三种细胞系：U373（星形胶质细胞），U937（单核细胞）和Daudi（B淋巴细胞）。使用抗针对α3，α4，α7，α9，β2和β4n9，β2和β4α9，β2和β4naChr亚基的抗体的荧光共聚焦显微镜。示出了研究的细胞在质膜上表达了不同的NACHR亚基，表明所有人都存在α7nachrs的存在U373细胞α3β4NACHR的细胞和U937细胞上的α4β2/α4β4NACHR。除了暴露在表面上的NACHRS之外，所有细胞均包含相当大的细胞内α3-和α7的NACHR。 α3-，α7-和β4特异性抗体的结合部分地与线粒体外膜旋转杂交酶特异性抗体的抗体部分重叠。 α7特异性抗体的结合也与有肌肉瘤绿色的抗体相结合，其与活性线粒体结合。所获得的数据表明，U373，U937和Daudi细胞中的细胞内α3β4和α7NACHRS的一部分位于线粒体上。该发现补充了我们之前对小鼠肝线粒体中α7NACHRS的观察结果，并揭示了胆碱能调节的新细胞内靶标。

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