title: Intracellular endothelin type B receptor-driven Ca2+ signal elicits nitric oxide production in endothelial cells.
abstract: Endothelin-1 exerts its actions via activation of ET(A) and ET(B) G(q/11) protein-coupled receptors, located in the plasmalemma, cytoplasm, and nucleus. Although the autocrine/paracrine nature of endothelin-1 signaling has been extensively studied, its intracrine role has been largely attributed to interaction with receptors located on nuclear membranes and the nucleoplasm. Because ET(B) receptors have been shown to be targeted to endolysosomes, we used intracellular microinjection and concurrent imaging methods to test their involvement in Ca(2+) signaling and subsequential NO production. We provide evidence that microinjected endothelin-1 produces a dose-dependent elevation in cytosolic calcium concentration in ET(B)-transfected cells and endothelial cells; this response is sensitive to ET(B) but not ET(A) receptor blockade. In endothelial cells, the endothelin-1-induced Ca(2+) response is abolished upon endolysosomal but not Golgi disruption. Moreover, the effect is prevented by inhibition of microautophagy and is sensitive to inhibitors of the phospholipase C and inositol 1,4,5-trisphosphate receptor. Furthermore, intracellular endothelin-1 increases nitric oxide via an ET(B)-dependent mechanism. Our results indicate for the first time that intracellular endothelin-1 activates endolysosomal ET(B) receptors and increase cytosolic Ca(2+) and nitric oxide production. Endothelin-1 acts in an intracrine fashion on endolysosomal ET(B) to induce nitric oxide formation, thus modulating endothelial function.
authors.affiliation: Department of Pharmacology, Temple University School of Medicine, Philadelphia, Pennsylvania 19140, USA.
authors.forename: Elena
authors.initials: E
authors.lastname: Deliu
authors.forename: G Cristina
authors.initials: GC
authors.lastname: Brailoiu
authors.forename: Karthik
authors.initials: K
authors.lastname: Mallilankaraman
authors.forename: Hong
authors.initials: H
authors.lastname: Wang
authors.forename: Muniswamy
authors.initials: M
authors.lastname: Madesh
authors.forename: Ashiwel S
authors.initials: AS
authors.lastname: Undieh
authors.forename: Walter J
authors.initials: WJ
authors.lastname: Koch
authors.forename: Eugen
authors.initials: E
authors.lastname: Brailoiu
mesh_terms: D000818:Animals; D001343:Autophagy; D002118:Calcium / Q000378:metabolism; D020013:Calcium Signaling; D002467:Cell Nucleus / Q000378:metabolism; D002478:Cells, Cultured; D003600:Cytosol / Q000378:metabolism; D011992:Endosomes / Q000378:metabolism; D042783:Endothelial Cells / Q000378:metabolism*; D019332:Endothelin-1 / Q000378:metabolism*; D008247:Lysosomes / Q000378:metabolism; D008833:Microcirculation; D008954:Models, Biological; D009569:Nitric Oxide / Q000378:metabolism*; D010084:Oxidation-Reduction; D051381:Rats; D044023:Receptor, Endothelin B / Q000378:metabolism*; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism
chemical_list: D019332:Endothelin-1; D044023:Receptor, Endothelin B; D043562:Receptors, G-Protein-Coupled; D009569:Nitric Oxide; D002118:Calcium