Saulière Aude, Gaibelet Gérald, Millot Claire, Mazères Serge, Lopez André, Salomé Laurence
Institut de Pharmacologie et Biologie Structurale, UMR CNRS 5089, 205, route de Narbonne, 31077 TOULOUSE cedex, France.
FEBS Lett. 2006 Oct 2;580(22):5227-31. doi: 10.1016/j.febslet.2006.08.056. Epub 2006 Sep 5.
Previous single-molecule studies have shown a long-term diffusion superimposed to a short-term confinement of the human mu opioid (hMOP) receptors at the surface of heterologous cells. However, additional ensemble average measurements are required to reach a complete understanding of the undergoing process. Here, we analyse, by fluorescence recovery after photobleaching measurements, the lateral diffusion of fully functional T7-EGFP-hMOP receptors in neuroblastoma SH-SY5Y cells naturally expressing a low level of the wild-type receptor. Experiments carried out at variable observation radii demonstrate the restriction of the receptors diffusion to sub-micrometer sized domains. Furthermore, consistently with the long-term single-molecule data, the domains are found permeable.
先前的单分子研究表明,在异源细胞表面,人类μ阿片受体(hMOP)存在叠加于短期限制之上的长期扩散现象。然而,需要进行额外的总体平均测量,才能全面了解这一过程。在此,我们通过光漂白后荧光恢复测量,分析了在天然表达低水平野生型受体的神经母细胞瘤SH-SY5Y细胞中,功能完全正常的T7-EGFP-hMOP受体的横向扩散。在可变观察半径下进行的实验表明,受体扩散被限制在亚微米大小的区域内。此外,与长期单分子数据一致,这些区域是可渗透的。