Co-localization of mu and delta opioid receptors on SK-N-SH cells detected by fluorescence microscopy using labeled anti-idiotypic antibodies.
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.
Agarwal
D
D
Department of Biochemistry, University of Connecticut Health Center, Farmington 06030.
Glasel
J A
JA
D000888:Antibodies, Anti-Idiotypic; D001667:Binding, Competitive; D020875:Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; D020881:Enkephalin, D-Penicillamine (2,5)-; D004745:Enkephalins / Q000378:metabolism; D006801:Humans; D008856:Microscopy, Fluorescence; D009447:Neuroblastoma / Q000648:ultrastructure; D011957:Receptors, Opioid / Q000145:classification; D017465:Receptors, Opioid, delta / Q000032:analysis* / Q000378:metabolism; D017450:Receptors, Opioid, mu / Q000032:analysis* / Q000378:metabolism; D014407:Tumor Cells, Cultured
D000888:Antibodies, Anti-Idiotypic; D004745:Enkephalins; D011957:Receptors, Opioid; D017465:Receptors, Opioid, delta; D017450:Receptors, Opioid, mu; D020875:Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; D020881:Enkephalin, D-Penicillamine (2,5)-