Endothelin inhibits vasopressin action in rat inner medullary collecting duct via the ETB receptor.
We characterized the endothelin (ET) receptor subtype responsible for the inhibition of vasopressin (AVP)-induced increases in osmotic water permeability (Pf) and cAMP accumulation in rat inner medullary collecting ducts (IMCD). ET-1 (10 nM) produced a rapid and transient decrease in AVP-stimulated Pf from 1241 +/- 112 to 224 +/- 38 microns/sec. At the same concentration (10 nM), the selective ETB receptor agonist sarafotoxin 6c (S6c) produced the same degree of inhibition with a time course identical to that of ET-1. Exposure of IMCDs to the ETA-selective antagonist BQ123 (100 nM) had no effect on ET-1-induced inhibition of AVP-dependent Pf. In suspensions of IMCD cells, ET-1, ET-3 or S6c produced concentration-dependent inhibition of AVP-stimulated cAMP accumulation to the same extent and with similar potencies (IC50 = 10-30 nM). BQ123 (1 nM to 10 microM) had no effect on ET-1-induced inhibition of AVP-stimulated cAMP formation. Saturation binding experiments with radiolabeled ET-1 and the selective ETB agonist IRL1620 and competition binding studies with selective ETA and ETB receptor ligands demonstrated that > or = 80% of the ET-1 binding sites in IMCD membranes were of the ETB subtype. Therefore, results from functional, biochemical and binding studies suggest that the ETB receptor is the ET receptor subtype that inhibits AVP action in the rat IMCD.
Edwards
R M
RM
Department of Renal Pharmacology, SmithKline Beecham Pharmaceuticals, King of Prussia, PA.
Stack
E J
EJ
Pullen
M
M
Nambi
P
P
D000818:Animals; D001127:Arginine Vasopressin / Q000037:antagonists & inhibitors*; D002463:Cell Membrane Permeability; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000378:metabolism; D065128:Endothelin Receptor Antagonists; D016232:Endothelins / Q000378:metabolism / Q000494:pharmacology*; D066298:In Vitro Techniques; D007457:Iodine Radioisotopes; D007679:Kidney Medulla / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D007685:Kidney Tubules, Collecting / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D007700:Kinetics; D008297:Male; D009997:Osmotic Pressure; D010446:Peptide Fragments / Q000378:metabolism; D010477:Perfusion; D051381:Rats; D017207:Rats, Sprague-Dawley; D017466:Receptors, Endothelin / Q000502:physiology; D014867:Water / Q000378:metabolism
D065128:Endothelin Receptor Antagonists; D016232:Endothelins; D007457:Iodine Radioisotopes; D010446:Peptide Fragments; D017466:Receptors, Endothelin; D014867:Water; D001127:Arginine Vasopressin; C074308:sovateltide; D000242:Cyclic AMP