title: Regulation of prostaglandin EP1 and EP4 receptor signaling by carrier-mediated ligand reuptake.
abstract: After synthesis and release from cells, prostaglandin E2 (PGE2) undergoes reuptake by the prostaglandin transporter (PGT), followed by cytoplasmic oxidation. Although genetic inactivation of PGT in mice and humans results in distinctive phenotypes, and although experiments in localized environments show that manipulating PGT alters downstream cellular events, a direct mechanistic link between PGT activity and PGE2 (EP) receptor activation has not been made. Toward this end, we created two reconstituted systems to examine the effect of PGT expression on PGE2 signaling via two of its receptors (EP1 and EP4). In human embryonic kidney cells engineered to express the EP1 receptor, exogenous PGE2 induced a dose-dependent increase in cytoplasmic Ca(2+). When PGT was expressed at the plasma membrane, the PGE2 dose-response curve was right-shifted, consistent with reduction in cell surface PGE2 availability; a potent PGT inhibitor acutely reversed this shift. When bradykinin was used to induce endogenous PGE2 release, PGT expression similarly induced a reduction in Ca(2+) responses. In separate experiments using Madin-Darby Canine Kidney cells engineered to express the PGE2 receptor EP4, bradykinin again induced autocrine PGE2 signaling, as judged by an abrupt increase in intracellular cAMP. As in the EP1 experiments, expression of PGT at the plasma membrane caused a reduction in bradykinin-induced cAMP accumulation. Pharmacological concentrations of exogenous PGE2 induced EP4 receptor desensitization, an effect that was mitigated by PGT. Thus, at an autocrine/paracrine level, plasma membrane PGT regulates PGE2 signaling by decreasing ligand availability at cell surface receptors.
authors.affiliation: Department of Medicine, Albert Einstein College of Medicine Bronx, New York, 10461.
authors.forename: Yuling
authors.initials: Y
authors.lastname: Chi
authors.affiliation: Department of Urology, Albert Einstein College of Medicine Bronx, New York, 10461 ; Department of Neuroscience, Albert Einstein College of Medicine Bronx, New York, 10461.
authors.forename: Sylvia O
authors.initials: SO
authors.lastname: Suadicani
authors.affiliation: Department of Medicine, Albert Einstein College of Medicine Bronx, New York, 10461 ; Department of Physiology & Biophysics, Albert Einstein College of Medicine Bronx, New York, 10461.
authors.forename: Victor L
authors.initials: VL
authors.lastname: Schuster
keywords: Eicosanoids; G-protein-coupled receptors; SLCO2A1; prostaglandins; signal transduction