title: [125I]-PD151242: a selective ligand for endothelin ETA receptors in human kidney which localizes to renal vasculature.
abstract: 1. The linear tetrapeptide radioligand, [125I]-PD151242 was used to characterize ETA receptors in human kidney which is an ETB-rich tissue. Saturation binding assays with [125I]-PD151242 revealed a single population of high affinity endothelin receptors: KD = 0.75 +/- 0.07 nM and Bmax = 48.4 +/- 1.6 fmol mg-1 protein (n = 3 individuals +/- s.e.mean). Hill slopes were close to unity and a one site fit was preferred to a two site model. 2. ETA-receptor-selective ligands competed for [125I]-PD151242 binding with sub-nanomolar affinity: BQ123 KD = 0.43 +/- 0.10 nM, Bmax = 46.6 +/- 7.9 fmol mg-1 protein; FR139317, KD = 0.37 +/- 0.06 nM, Bmax = 39.5 +/- 6.5 fmol mg-1 protein (n = 3 individuals +/- s.e.mean). In each case, monophasic inhibition curves were obtained and a one site fit was preferred to a two site model. The ETB-selective agonist, BQ3020 at the highest concentration tested (10 microM) inhibited binding by only 50%. The non-selective RO462005 competed for the binding of [125I]-PD151242: KD = 1.31 +/- 1.38 microM, Bmax = 33.0 +/- 9.7 fmol mg-1 protein. Endothelin-2 and sarafotoxin S6B inhibited [125I]-PD151242 binding to renal tissue whereas ET-3 and sarafotoxin S6C were less effective. Non-endothelin and non-sarafotoxin peptides did not compete. 3. No degradation of [125I]-PD151242 was detected following incubation of the ligand with renal tissue under the conditions of the binding assay. 4. Polymerase chain reaction products corresponding to the expected size for mRNA encoding ETA and ETB receptor sub-types were detected in cortex and medulla in each of the five individuals examined.5. Autoradiographical studies showed that ETA receptors visualised with ['25I]-PD151242 were mainly localized to blood vessels including interlobular and arcuate arteries, arterioles and adjacent arcuate veins. ETB receptors localized with ['251]-BQ3020 were concentrated in the medulla and the density of binding to vessels was low.6. These data suggest [251I]-PDl51242 is selective for ETA receptors in human kidney and this sub-type is mainly localized to the renal vasculature. The results provide further evidence that the human vasculature mainly expresses the ETA receptor.
authors.affiliation: Clinical Pharmacology Unit, University of Cambridge, Addenbrooke's Hospital.
authors.forename: A P
authors.initials: AP
authors.lastname: Davenport
authors.forename: R E
authors.initials: RE
authors.lastname: Kuc
authors.forename: S L
authors.initials: SL
authors.lastname: Hoskins
authors.forename: F E
authors.initials: FE
authors.lastname: Karet
authors.forename: F
authors.initials: F
authors.lastname: Fitzgerald
mesh_terms: D000328:Adult; D000368:Aged; D000595:Amino Acid Sequence; D001345:Autoradiography; D001381:Azepines; D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D004587:Electrophoresis, Agar Gel; D065128:Endothelin Receptor Antagonists; D006801:Humans; D066298:In Vitro Techniques; D007668:Kidney / Q000187:drug effects / Q000378:metabolism*; D007700:Kinetics; D008024:Ligands; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D009842:Oligopeptides; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism; D017466:Receptors, Endothelin / Q000819:agonists / Q000187:drug effects*; D012079:Renal Circulation / Q000187:drug effects*
chemical_list: D001381:Azepines; D065128:Endothelin Receptor Antagonists; D008024:Ligands; D009842:Oligopeptides; D012333:RNA, Messenger; D017466:Receptors, Endothelin; C087850:PD 151242; D012194:RNA-Directed DNA Polymerase