title: Typical endothelin ETA receptors mediate atypical endothelin-1-induced contractions in sheep isolated tracheal smooth muscle.
abstract: Contraction of vascular and nonvascular smooth muscle induced by the endothelin/sarafotoxin family of peptides frequently does not readily fit into the current classification criteria for ETA and ETB receptors, raising the possibility of additional atypical receptors. In the current study, isometric tension recording and radioligand binding techniques were used to characterize the ETA receptor population in sheep isolated tracheal smooth muscle. Endothelin-1 and sarafotoxin S6b induced similar concentration-dependent contractions, although endothelin-1 was 2.6-fold more potent (P <.05, n = 15-18). The ETA receptor-selective antagonists BQ-123 and FR139317 caused concentration-dependent inhibition of the contractions induced by endothelin-1 and sarafotoxin S6b, but both antagonists were significantly less potent in inhibiting contractions induced by endothelin-1 than sarafotoxin S6b. For example, 0.03 microM FR139317 shifted the endothelin-1 and sarafotoxin S6b concentration-effect curves to the right by 1.8- and 8.3-fold, respectively (P <.01, n = 6-8). Although the observed agonist dependence of antagonist potency may indicate the presence of atypical ETA receptors, competition binding studies using 125I-endothelin-1 and 125-I-sarafotoxin S6b identified only a single population of BQ-123- and sarafotoxin S6b-sensitive ETA receptors. Additional association-, dissociation-, and saturation-binding studies revealed that 125I-endothelin-1 binding to these ETA receptors was pseudoirreversible, whereas 125I-sarafotoxin S6b binding was readily reversible. Thus, marked differences in the kinetic profiles of ETA receptor binding to endothelin-1, sarafotoxin S6b, and BQ-123, rather than the existence of another ETA receptor subtype, may explain the stark agonist dependence of antagonist potency observed in contractile studies.
authors.affiliation: Department of Pharmacology, University of Western Australia, Nedlands, Australia. phenry@receptor.pharm.uwa.edu.au
authors.forename: P J
authors.initials: PJ
authors.lastname: Henry
authors.forename: S H
authors.initials: SH
authors.lastname: King
mesh_terms: D000818:Animals; D001381:Azepines / Q000494:pharmacology; D001667:Binding, Competitive; D065128:Endothelin Receptor Antagonists; D019332:Endothelin-1 / Q000494:pharmacology*; D066298:In Vitro Techniques; D007211:Indoles / Q000494:pharmacology; D007700:Kinetics; D009119:Muscle Contraction / Q000187:drug effects / Q000502:physiology*; D009130:Muscle, Smooth / Q000187:drug effects / Q000502:physiology*; D010456:Peptides, Cyclic / Q000494:pharmacology; D011869:Radioligand Assay; D044022:Receptor, Endothelin A; D017466:Receptors, Endothelin / Q000378:metabolism / Q000502:physiology*; D012756:Sheep; D014132:Trachea / Q000187:drug effects / Q000502:physiology*; D014662:Vasoconstrictor Agents / Q000494:pharmacology; D014757:Viper Venoms / Q000494:pharmacology
chemical_list: D001381:Azepines; D065128:Endothelin Receptor Antagonists; D019332:Endothelin-1; D007211:Indoles; D010456:Peptides, Cyclic; D044022:Receptor, Endothelin A; D017466:Receptors, Endothelin; D014662:Vasoconstrictor Agents; D014757:Viper Venoms; C057354:sarafotoxins s6; C079574:FR 139317; C072247:cyclo(Trp-Asp-Pro-Val-Leu)