Agarwal D, Glasel J A
Department of Biochemistry, University of Connecticut Health Center, Farmington 06030.
Life Sci. 1993;52(18):PL193-8. doi: 10.1016/0024-3205(93)90117-l.
Selective fluorescence labeling of opioid receptor subclasses on SK-N-SH cultured cells has been accomplished using labeled polyclonal anti-idiotypic antibodies along with subclass-selective opioid agonists (DPDPE, delta-selective; DAMGO, mu-selective) as blocking reagents. Labeling of the cells was examined using conventional fluorescence microscopy. Co-localization of mu- and delta-opioid receptors on SK-N-SH cells has been studied by double labeling fluorescence experiments. In agreement with our own, and other workers', previous observations on NG108-15 cells, a subpopulation of viable cells in asynchronous cultures are labeled. Among those SK-N-SH cells that are labeled, both subclasses of receptors are seen. On the basis of sequential blocking experiments we interpret our combined results to be consistent with a model where mu- and delta- binding sites reside on different subunits of a multimeric complex.
利用标记的多克隆抗独特型抗体以及亚类选择性阿片样物质激动剂（DPDPE，δ选择性；DAMGO，μ选择性）作为封闭试剂，已在SK-N-SH培养细胞上实现了阿片受体亚类的选择性荧光标记。使用传统荧光显微镜检查细胞的标记情况。通过双标记荧光实验研究了SK-N-SH细胞上μ和δ阿片受体的共定位。与我们自己以及其他研究人员先前对NG108-15细胞的观察结果一致，异步培养的活细胞亚群被标记。在那些被标记的SK-N-SH细胞中，可以看到两种受体亚类。基于顺序封闭实验，我们将综合结果解释为与一种模型一致，即μ和δ结合位点位于多聚体复合物的不同亚基上。
