title: Heterogeneous platelet-activating factor (PAF) receptors and calcium increase in platelets and macrophages.
abstract: We used the increase in cytosolic Ca2+ levels, [Ca2+]i, as a way to characterize PAF (platelet-activating factor, 1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine) receptors in human platelets and rat and human macrophages. [Ca2+] was measured by means of the fluorescent probe fura-2/acetoxymethylester. PAF recognized heterogeneous receptors in human macrophages only (curve slope <1). The PAF antagonist SCH 37370 (1-acetyl-4(8-chloro-5,6-dihydro-11H-benzo[5.6]cyclohepta[1,2-b]pyridine -11-ylidine)piperidine) abolished [Ca2+]i elevation in human platelets, while in rat and human macrophages the maximal inhibition was 76% and 85%, respectively. On the contrary, the antagonist WEB 2086 (3-[4-(2-chlorophenyl)-9-methyl-6Hthieno[3,2-f] [1,2,4]triazolo-[4,3-a] [1,4]-diazepin-2-yl]-1-(4-morpholiny)-1-propanon, apafant) totally inhibited the effect of PAF in both platelets and macrophages. The WEB 2086 concentration-response curves had a slope <1 in the three cell types, indicating interaction with heterogeneous receptors. Accordingly, 3H-WEB 2086 bound to two different classes of sites. Both phases of [Ca2+]i elevation (influx or release) were equally affected by the antagonists. These data support the notions that: 1) PAF receptors are heterogeneous; 2) the two antagonists have a different selectivity toward the receptor subtypes: WEB 2086 recognizes two different receptors both in platelets and in macrophages, while SCH 37370 does not discriminate between receptor subtypes in platelets, and only interacts with one subtype in macrophages; and 3) both SCH 37370 and WEB 2086 display different potencies in rat and human macrophages.
authors.affiliation: Laboratory of Molecular Pharmacology, Institute of Pharmacological Sciences, and E. Grossi Paoletti Center, University of Milan, Italy.
authors.forename: C
authors.initials: C
authors.lastname: Centemeri
authors.forename: S
authors.initials: S
authors.lastname: Colli
authors.forename: D
authors.initials: D
authors.lastname: Tosarello
authors.forename: P
authors.initials: P
authors.lastname: Ciceri
authors.forename: S
authors.initials: S
authors.lastname: Nicosia
mesh_terms: D000818:Animals; D001381:Azepines / Q000494:pharmacology; D001792:Blood Platelets / Q000378:metabolism*; D002118:Calcium / Q000097:blood*; D003600:Cytosol / Q000378:metabolism; D006801:Humans; D017336:Loratadine / Q000031:analogs & derivatives; D008264:Macrophages / Q000378:metabolism*; D008297:Male; D010880:Piperidines / Q000494:pharmacology; D010980:Platelet Membrane Glycoproteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011869:Radioligand Assay; D051381:Rats; D017207:Rats, Sprague-Dawley; D011956:Receptors, Cell Surface; D043562:Receptors, G-Protein-Coupled; D014230:Triazoles / Q000494:pharmacology
chemical_list: D001381:Azepines; D010880:Piperidines; D010980:Platelet Membrane Glycoproteins; D011956:Receptors, Cell Surface; D043562:Receptors, G-Protein-Coupled; D014230:Triazoles; C039351:platelet activating factor receptor; C052518:WEB 2086; D017336:Loratadine; C063524:N-acetyldesloratadine; D002118:Calcium