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J Biol Chem, Vol. 274, Issue 22, 15719-15724, May 28, 1999

Localization of Functional Prostaglandin E2 Receptors EP3 and EP4 in the Nuclear Envelope

Mousumi BhattacharyaDagger , Krishna Peri, Alfredo Ribeiro-da-SilvaDagger , Guillermina AlmazanDagger , Hitoshi Shichiparallel , Xin Hou, Daya R. VarmaDagger , and Sylvain Chemtob**

From the Dagger  Department of Pharmacology & Therapeutics, McGill University, Montreal, PQ, Canada, H3G 1Y6, the ** Departments of Pediatrics, Ophthalmology and Pharmacology, and the  Research Center of Hôpital Ste. Justine, University of Montreal, Montreal, PQ, Canada H3T 1C5, and the parallel  Department of Ophthalmology, Wayne State University School of Medicine, Detroit, Michigan 48201

The effects of prostaglandin E2 are thought to be mediated via G protein-coupled plasma membrane receptors, termed EP. However recent data implied that prostanoids may also act intracellularly. We investigated if the ubiquitous EP3 and the EP4 receptors are localized in nuclear membranes. Radioligand binding studies on isolated nuclear membrane fractions of neonatal porcine brain and adult rat liver revealed the presence of EP3 and EP4. A perinuclear localization of EP3alpha and EP4 receptors was visualized by indirect immunocytofluorescence and confocal microscopy in porcine cerebral microvascular endothelial cells and in transfected HEK 293 cells that stably overexpress these receptors. Immunoelectron microscopy clearly revealed EP3alpha and EP4 receptors localization in the nuclear envelope of endothelial cells; this is the first demonstration of the nuclear localization of these receptors. Data also reveal that nuclear EP receptors are functional as they affect transcription of genes such as inducible nitric-oxide synthase and intranuclear calcium transients; this appears to involve pertussis toxin-sensitive G proteins. These results define a possible molecular mechanism of action of nuclear EP3 receptors.

Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.


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