PMID: 9435191Jan 22, 1998Paper

Detection of kappa opioid receptors on mouse thymocyte phenotypic subpopulations as assessed by flow cytometry

The Journal of Pharmacology and Experimental Therapeutics
T A Ignatowski, J M Bidlack

Abstract

Recent studies have shown kappa opioid receptor labeling on the R1EGO thymoma cell line by indirect immunofluorescence and flow cytometric analysis. The present study used a fluorescein-labeled arylacetamide (FITC-AA), a kappa opioid ligand, in conjunction with biotin-conjugated anti-fluorescein IgG and extravidin-R-phycoerythrin (PE), along with double-labeling with antibodies against specific immune cell surface markers to determine which subpopulation(s) of thymocytes express the kappa opioid receptor. Thymocytes, isolated from 6- to 8-week-old C57BL/6ByJ mice, incubated with FITC-AA followed by the PE amplification procedure, demonstrated labeling of the kappa opioid receptor. This labeling was inhibited 55 +/- 4% above background by excess nor-binaltorphimine (nor-BNI), a kappa selective antagonist. This kappa opioid receptor positive population consisted of 58 +/- 2% of all gated thymocytes. Phenotypic characterization determined that not only were 64 +/- 3% of the gated thymocytes CD4+/kappa opioid receptor positive, but 60 +/- 1% of all thymocytes were CD8+/kappa opioid receptor positive. Two subpopulations of CD3+ thymocytes, consisting of both mature and immature cells, also displayed labeling for the kappa opioid rec...Continue Reading

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