Rapid desensitization and slow recovery of the cyclic AMP response mediated by histamine H2 receptors in the U937 cell line

doi:10.1016/S0006-2952(00)00295-1

Biochemical Pharmacology

Volume 60, Issue 2, 15 July 2000, Pages 159-166

https://doi.org/10.1016/S0006-2952(00)00295-1 Get rights and content

Abstract

The present study focused on the desensitization process of the H₂ receptor in U937 cells and the recovery of the cyclic AMP (cAMP) response. Treatment of U937 leukemic cells with the H₂ histamine receptor agonists (±)-N¹-[3-(3,4-difluorophenyl)-3-(pyridin-2-yl)propyl]-N²-[3-(1H-imidazol-4-yl)propyl]guanidine (BU-E-75) and amthamine produced a rapid desensitization characterized by decreased cAMP production (T_1/2 = 20 min). Pretreatment with 10 μM BU-E-75 did not induce modifications in the responses to prostaglandin E₂, isoproterenol, or forskolin. H₂ receptor desensitization was not affected by protein kinase A and C inhibitors, but was reduced drastically by Zn²⁺ and heparin, known to act as inhibitors of G protein-coupled receptor kinases. Recovery studies of the cAMP response showed that cAMP levels reached 50% of the initial values within 5 hr. Furthermore, desensitization produced an important decrease in the basal level of this cyclic nucleotide. The minimal value was observed 12 hr later, and corresponded to approximately 1.3% of the initial basal level (7.5 vs 0.1 pmol/10⁶ cells). This result could be explained by an increase in phosphodiesterase activity following 10 μM BU-E-75 treatment. When cells were exposed for 2 hr to an H₂ agonist, binding assays showed no modification in the number of H₂ receptors; internalization began just after 8 hr. Although the initial desensitization seems to involve G protein-coupled receptor kinases, results indicate that additional mechanisms of regulation were triggered by the H₂ agonists.

Section snippets

Chemicals

Histamine dihydrochloride, forskolin, PGE₂, IBMX, isoproterenol, cAMP, 1-(5-isoquinolinylsulphonyl)-2-methylpiperazine (H7), famotidine, and mepyramine were obtained from the Sigma Chemical Co. Amthamine, tiotidine, and N-(2-aminoethyl)-5-isoquinolinesulphonamide (H9) were purchased from Tocris Cookson Inc. 2′-O-Anthraniloyl cAMP was purchased from GIBCO BRL. Dimaprit, (±)-N¹-[3-(4-fluorophenyl)-3-(pyridin-2-yl)propyl]-N²-[3-(1H-imidazol-4-yl)propyl]guanidine (arpromidine), and (±)-N¹

cAMP production by H₂ agonists in U937 cells

The ability of U937 cells to produce cAMP when stimulated with histamine or dimaprit was similar to that reported by other authors [13]. Similar results were obtained with other H₂ agonists such as BU-E-75, arpromidine [21], and amthamine (Table 1).

Histamine H₂ receptor desensitization

H₂ receptor desensitization assays were performed with specific H₂ agonists to avoid interactions between the signaling pathways of both receptor subtypes, as previously described [22].

After pretreatment ranging from 1 min to 3 hr with a given H₂

Discussion

The U937 cell line is derived from the pleural fluid of a patient with diffuse histiocytic lymphoma [23]. The presence of H₂ receptors in this cell line has been described, showing a classic pharmacological profile similar to the one described for the guinea pig right atrium [13]. For this reason, these cells represent an interesting model for studying the signal transduction pathway for H₂ receptors. Recent studies indicate a rapid desensitization of the H₂ receptor by 100 μM histamine [13].

Acknowledgements

This study was supported by grants from the Antorchas Foundation (A-13218/1–000026), the University of Buenos Aires (JB10), and CONICET-FONCYT, Argentina (PICT-1598; 4406).

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Molecular and cellular pharmacologyRapid desensitization and slow recovery of the cyclic AMP response mediated by histamine H2 receptors in the U937 cell line

Abstract

Section snippets

Chemicals

cAMP production by H2 agonists in U937 cells

Histamine H2 receptor desensitization

Discussion

Acknowledgements

J Biol Chem

Biochim Biophys Acta

Biochem Biophys Res Commun

Eur J Pharmacol

Eur J Pharmacol

Biochem Pharmacol

Methods Enzymol

J Biol Chem

Trends Pharmacol Sci

Biochem Biophys Res Commun

Pharmacol Ther

Pharmacol Ther

J Biol Chem

Exp Cell Res

International Union of Pharmacology. XIIIClassification of histamine receptors

Pharmacol Rev

Molecular cloning of a gene encoding the histamine H2 receptor

Proc Natl Acad Sci USA

Model systems for the study of seven-transmembrane segment-receptors

Annu Rev Biochem

G proteins of the Gq family couple the H2 histamine receptor to phospholipase C

Mol Endocrinol

Turning of the signalMechanisms that attenuate signaling by G protein-coupled receptors

Am J Physiol

Molecular and cellular pharmacology
Rapid desensitization and slow recovery of the cyclic AMP response mediated by histamine H₂ receptors in the U937 cell line

cAMP production by H₂ agonists in U937 cells

Histamine H₂ receptor desensitization

Molecular cloning of a gene encoding the histamine H₂ receptor

G proteins of the G_q family couple the H₂ histamine receptor to phospholipase C