Werner, Kristin and Kaelble, Solveig and Wolter, Sabine and Schneider, Erich H. and Buschauer, Armin and Neumann, Detlef and Seifert, Roland (2015) Flow cytometric analysis with a fluorescently labeled formyl peptide receptor ligand as a new method to study the pharmacological profile of the histamine H-2 receptor. NAUNYN-SCHMIEDEBERGS ARCHIVES OF PHARMACOLOGY, 388 (10). pp. 1039-1052. ISSN 0028-1298, 1432-1912
Full text not available from this repository. (Request a copy)Abstract
The histamine H-2 receptor (H2R) is a G(s) protein-coupled receptor. Its activation leads to increases in the second messenger adenosine-3',5'-cyclic monophosphate (cAMP). Presently, several systems are established to characterize the pharmacological profile of the H2R, mostly requiring radioactive material, animal models, or human blood cells. This prompted us to establish a flow cytometric analysis with a fluorescently labeled formyl peptide receptor (FPR) ligand in order to investigate the H2R functionally and pharmacologically. First, we stimulated U937 promonocytes, which mature in a cAMP-dependent fashion upon H2R activation, with histamine (HA) or selective H2R agonists and measured increases in cAMP concentrations by mass spectrometry. Next, indicative for the maturation of U937 promonocytes, we assessed the FPR expression upon incubation with HA or H2R agonists. FPR expression was measured either indirectly by formyl peptide-induced changes in intracellular calcium concentrations ([Ca2+](i)) or directly with the fluorescein-labeled FPR ligand fNleLFNleYK-Fl. HA and H2R agonists concentration-dependently induced FPR expression, and potencies and efficacies of fMLP-induced increases in [Ca2+](i) and FPR density correlated linearly. Accordingly, flow cytometric analysis of FPR expression constitutes a simple, inexpensive, sensitive, and reliable method to characterize the H2R pharmacologically. Furthermore, we evaluated FPR expression at the mRNA level. Generally, quantitative real-time polymerase chain reaction confirmed functional data. Additionally, our study supports the concept of functional selectivity of the H2R, since we observed dissociations in the efficacies of HA and H2R agonists in cAMP accumulation and FPR expression.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | FUNCTIONAL SELECTIVITY; CONSTITUTIVE ACTIVITY; GUINEA-PIG; HL-60; DIFFERENTIATION; EXPRESSION; DESENSITIZATION; CONFORMATIONS; U-937; PCR; Histamine H-2 receptor; Formyl peptide receptor; Functional selectivity; U937 promonocytes |
| Subjects: | 500 Science > 540 Chemistry & allied sciences |
| Divisions: | Chemistry and Pharmacy > Institute of Pharmacy > Pharmaceutical/Medicinal Chemistry II (Prof. Buschauer) |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 07 Jun 2019 06:28 |
| Last Modified: | 07 Jun 2019 06:29 |
| URI: | https://pred.uni-regensburg.de/id/eprint/4798 |
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