Characterization of the prostaglandin E receptor expressed on a cultured mast cell line, BNu-2cl3.
Interleukin 3-dependent BNu-2cl3 mast cells, mucosal type-like mast cells, exhibited a specific high-affinity binding site for [3H]prostaglandin (PG) E2. The binding was completely displaced by M&B 28767, an EP3-selective agonist, but not by EP1- or EP2-selective ligands, indicating that the PGE2 binding site is of the EP3 subtype PGE receptor. Whereas the EP3 subtype is presumed to be coupled to inhibition of adenylate cyclase in various tissues and cells, in BNu-2cl3 cells PGE2 had no ability to inhibit adenylate cyclase activity, while it induced concentration-dependent stimulation of phosphoinositide metabolism and caused an increase in the intracellular free Ca2+ concentration in a pertussis toxin-sensitive manner. PGE2 by itself did not evoke histamine release from the cells, but it markedly stimulated histamine release in concert with ionomycin, a Ca2+ ionophore. The PGE2-stimulated release was also completely blocked by pertussis toxin. Thus, the PGE receptor expressed on BNu-2cl3 mast cells is of the EP3 subtype and is linked to phosphoinositide metabolism via a pertussis toxin-sensitive G protein, and this activation leads to histamine release.
Nishigaki
N
N
Department of Physiological Chemistry, Faculty of Pharmaceutical Sciences, Kyoto University, Japan.
Negishi
M
M
Sugimoto
Y
Y
Namba
T
T
Narumiya
S
S
Ichikawa
A
A
D037361:Adenylate Cyclase Toxin; D000818:Animals; D001665:Binding Sites; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D006636:Histamine Release; D007377:Interleukin-3 / Q000494:pharmacology; D015759:Ionomycin / Q000494:pharmacology; D008407:Mast Cells / Q000378:metabolism*; D051379:Mice; D037342:Pertussis Toxin; D010716:Phosphatidylinositols / Q000378:metabolism; D011982:Receptors, Prostaglandin / Q000378:metabolism*; D018078:Receptors, Prostaglandin E; D015398:Signal Transduction; D014316:Tritium; D010566:Virulence Factors, Bordetella / Q000494:pharmacology
D037361:Adenylate Cyclase Toxin; D007377:Interleukin-3; D010716:Phosphatidylinositols; D011982:Receptors, Prostaglandin; D018078:Receptors, Prostaglandin E; D010566:Virulence Factors, Bordetella; D014316:Tritium; D015759:Ionomycin; D037342:Pertussis Toxin