Both endothelin-A and endothelin-B receptors are present on adult rat cardiac ventricular myocytes.
Endothelin-A (ET(A)) and endothelin-B (ET(B)) receptors have been demonstrated in intact heart and cardiac membranes. ET(A) receptors have been demonstrated on adult ventricular myocytes. The aim of the present study was to determine the presence of ET(B) and the relative contribution of this receptor subtype to total endothelin-1 (ET-1) binding on adult ventricular myocytes. Saturation binding experiments indicated that ET-1 bound to a single population of receptors (Kd = 0.52 +/- 0.13 nM, n = 4) with an apparent maximum binding (Bmax) of 2.10 +/- 0.25 sites (x 10(5))/cell (n = 4). Competition experiments using 40 pM [125I]ET-1 and nonradioactive ET-1 revealed a Ki of 660 +/- 71 pM (n = 10) and a Hill coefficient (nH) of 0.99 +/- 0.10 (n = 10). A selective ET(A) antagonist, BQ610, displaced 80% of the bound [125I]ET-1. No displacement was observed by concentrations of an ET(B)-selective antagonist, BQ788, up to 1.0 microM. However, in the presence of 1.0 microM BQ610, BQ788 inhibited the remaining [125I]ET-1 binding. Similarly, in the presence of 1.0 microM BQ788, BQ610 inhibited the remaining specific [125I]ET-1 binding. Binding of an ET(B1)-selective agonist, [125I]IRL-1620, confirmed the presence of ET(B). ET(B) bound to ET-1 irreversibly, whereas binding to ET(A) demonstrated both reversible and irreversible components, and BQ610 and BQ788 bound reversibly. Reducing the incubation temperature to 0 degrees C did not alter the irreversible component of ET-1 binding. Hence, both ET(A) and ET(B) receptors are present on intact adult rat ventricular myocytes, and the ratio of ET(A):ET(B) binding sites is 4:1. Both receptor subtypes bind to ET-1 by a two-step association involving the formation of a tight receptor-ligand complex; however, the kinetics of ET-1 binding to ET(A) versus ET(B) differ.
Allen
Bruce G
BG
Department of Medicine and Groupe de recherche sur le système nerveux autonome, Université de Montréal, Montréal, PQ H3C 3J7, Canada. allen@icm.umontreal.ca
Phuong
Luu Lien
LL
Farhat
Hala
H
Chevalier
Dominique
D
D000818:Animals; D001665:Binding Sites; D019332:Endothelin-1 / Q000493:pharmacokinetics*; D016232:Endothelins / Q000493:pharmacokinetics*; D006352:Heart Ventricles; D008297:Male; D032383:Myocytes, Cardiac / Q000737:chemistry*; D009842:Oligopeptides / Q000493:pharmacokinetics*; D010446:Peptide Fragments / Q000493:pharmacokinetics*; D010880:Piperidines / Q000493:pharmacokinetics*; D051381:Rats; D017207:Rats, Sprague-Dawley
D019332:Endothelin-1; D016232:Endothelins; D009842:Oligopeptides; D010446:Peptide Fragments; D010880:Piperidines; C074308:sovateltide; C087220:BQ 610; C086539:BQ 788