OATP2A1/SLCO2A1-mediated prostaglandin E2 loading into intracellular acidic compartments of macrophages contributes to exocytotic secretion

There is significant evidence that the inducible cyclooxygenase isoform (COX-2) regulates the pericellular concentration of PGE(2); however, the mechanism of the secretory process remains unclear. The present study, therefore, aimed to evaluate the role of prostaglandin transporter (OATP2A1) in PGE(2) secretion from macrophages. lmmunofluorescence staining for Oatp2a1 (Slco2a1) was primarily detected in cytoplasmic domains, and was partially co-localized with anti-PGE(2) antibody, LysoTracker (R), and anti-lysosome-associated membrane protein (Lamp) I antibody in murine macrophage-derived RAW264 cells and peritoneal macrophages (PMs). PGE(2) uptake by subcellular fraction containing light lysosomes was reduced significantly in the presence of an OATP inhibitor and in Slco2a1(+/-) PMs. Secretion of PGE(2) and lysosome-specific N-acetyl-beta-D-glucosaminidase was enhanced in activated macrophagic cells, and diminished significantly under the Ca2+-depleted condition. The amount of PGE(2) secreted from lipopolysaccharide-activated Slco2a1(-/-) PMs was significantly lower than that from PMs from wild type (WT) mice. Expression of Cox-2 and 15-hydroxyprostaglandin dehydrogenase (15-Pgdh) was unchanged between PMs from Slco2a1(-/-) and WT mice. These results suggest that OATP2A1 is involved in PGE(2)-loading into intracellular acidic compartments, including light lysosomes. Thus, OATP2A1 contributes to PGE(2) secretion by macrophages via exocytosis induced by Ca2+ influx, independently of PGE(2) synthesis and metabolism. (C) 2015 Elsevier Inc. All rights reserved.