title: Cell-specific differences in ET-1 system in adjacent layers of main pulmonary artery. A new source of ET-1.
abstract: Endothelin-1 (ET-1) is a potent vasoconstrictor that causes sustained constriction of the pulmonary artery and modulates normal vascular tone. Endothelial cells were thought to be the major source of ET-1, but recent studies show that vascular smooth muscle cells (SMCs) are also capable of its synthesis. We examined the ET-1 and endothelin-converting enzyme-1 (ECE-1) system in cells cultured from two adjacent layers, subendothelial (L1) and inner medial (L2), of normal sheep main pulmonary artery and the response of this system to exogenous ET-1 and transforming growth factor-beta1 (TGF-beta1). End points include assessment of preproET-1 (ppET-1) and ECE-1 gene coexpression, measurement of intracellular and released ET-1, and ECE-1 activity. RT-PCR analysis revealed that ppET-1 and ECE-1 transcripts were greater in L1 than in L2 cells. The L1 cells also synthesized (L1, 3.2 +/- 0.1; L2, 1.2 +/- 0.1 fmol/10(6) cells) and released (L1, 9.2 +/- 0.5; L2, 2.3 +/-0.1 fmol/ml) greater amounts of ET-1 than L2 cells. The L2 cells internalized exogenous ET-1 in a dose-dependent manner (EC(50) 8 nmol/l) and were more responsive to exogenous ET-1 than L1 cells, showing upregulation of both the ppET-1 and ECE genes. TGF-beta1 downregulated ET-1-stimulated ppET-1 and ECE-1 transcripts but only in L2 cells. In addition, L1 cells showed greater ECE-1 activity than L2 cells, and in both, the activity was sensitive to the metalloprotease inhibitor phosphoramidon. We conclude that the ET-1 system in L1 and L2 cells is distinct. The data suggest that the two cell types have diverse functions in the arterial wall; the L1 cells, like endothelial cells, provide a local source of ET-1; and since the L2 cells are more responsive to exogenous ET-1, they are likely to affect normal pulmonary vascular tone.
authors.affiliation: Department of Pathology, Center for Lung Research, Vanderbilt University Medical Center, Nashville, TN 37232-2650, USA.
authors.forename: E
authors.initials: E
authors.lastname: Tchekneva
authors.forename: M L
authors.initials: ML
authors.lastname: Lawrence
authors.forename: B
authors.initials: B
authors.lastname: Meyrick
mesh_terms: D000818:Animals; D016282:Aspartic Acid Endopeptidases / Q000235:genetics / Q000378:metabolism; D002478:Cells, Cultured; D019332:Endothelin-1 / Q000378:metabolism* / Q000494:pharmacology; D000072396:Endothelin-Converting Enzymes; D016232:Endothelins / Q000235:genetics; D015870:Gene Expression / Q000187:drug effects; D007425:Intracellular Membranes / Q000378:metabolism; D008666:Metalloendopeptidases; D008858:Microscopy, Phase-Contrast; D011498:Protein Precursors / Q000235:genetics; D011651:Pulmonary Artery / Q000166:cytology / Q000378:metabolism*; D012756:Sheep; D016212:Transforming Growth Factor beta / Q000494:pharmacology
chemical_list: D019332:Endothelin-1; D016232:Endothelins; D011498:Protein Precursors; D016212:Transforming Growth Factor beta; D016282:Aspartic Acid Endopeptidases; D008666:Metalloendopeptidases; D000072396:Endothelin-Converting Enzymes