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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：烟碱乙酰胆碱受体在人细胞系中的细胞内定位。

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摘要

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：先前我们证明了小鼠肝线粒体表达功能性α7烟碱乙酰胆碱受体（nAChRs）。本研究的目的是调查在非神经元人类细胞线粒体中是否存在nAChRs。流式细胞仪，细胞ELISA检测了三种细胞系：U373（星形细胞），U937（单核细胞）和Daudi（B淋巴细胞）。荧光共聚焦显微镜和抗α3，α4，α7，α9，β2和β4nAChR亚基胞外表位的抗体。研究表明，所研究的细胞在质膜上表达了不同的nAChR亚基集，表明所有α7nAChRs均存在U373细胞上的α3β4nAChRs和U937细胞上的α4β2/α4β4nAChRs。除了暴露在表面的nAChRs，所有细胞还包含大量的含α3和α7的nAChRs的细胞内池。 α3，α7和β4特异性抗体的结合与线粒体外膜转酶特异性抗体的结合部分重叠。 α7特异性抗体的结合与MitoTracker Green的结合也重叠，后者与活性线粒体结合。获得的数据表明，U373，U937和Daudi细胞中的一部分细胞内α3β4和α7nAChR位于线粒体上。这一发现补充了我们先前对小鼠肝线粒体中α7nAChRs的观察，并揭示了胆碱能调节的新细胞内靶标。

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