Functional analysis of natriuretic peptide receptors in the bladder of the toad, Bufo marinus.
This study aimed to localize and characterize natriuretic peptide binding sites in the urinary bladder of Bufo marinus and to then examine the effect of natriuretic peptides on the bladder vascular tone and water reabsorption in isolated perfused bladder preparations. Specific (125)I-rat atrial natriuretic peptide ((125)I-rANP) binding sites were present on blood vessels, muscle, and epithelium. In tissue sections and/or isolated membranes, the binding was completely displaced by frog ANP, rat ANP, and porcine C-type natriuretic peptide (CNP; membranes only). However, a reduction in binding was observed after incubation with (125)I-rANP and 1 microM of the natriuretic peptide receptor-C (NPR-C) ligand C-ANF, but residual binding remained suggesting the presence of two distinct binding sites. Electrophoresis of bladder membranes cross-linked to (125)I-rANP identified two bands at approximately 70 and 140 kDa that correspond to the monomeric mass of NPR-C and the guanylate cyclase receptors, respectively. Furthermore, the presence of natriuretic peptide receptor-A and NPR-C mRNA in the bladder was demonstrated with reverse transcription--polymerase chain reaction. In addition, rat ANP, frog ANP, and porcine CNP stimulated a significant increase in cGMP generation in bladder membrane preparations, which indicated the presence of guanylate cyclase-linked receptors. In perfused bladder preparations, arginine vasotocin increased perfusion pressure and water permeability. The infusion of frog ANP or porcine CNP failed to alter perfusion pressure or water reabsorption in the presence or absence of arginine vasotocin. This study identified a well-developed natriuretic peptide receptor system in the urinary bladder of B. marinus but the function of the receptors remains unclear.
Meier
Stuart K
SK
School of Biological and Chemical Sciences, Deakin University, Geelong, Victoria, 3217, Australia.
Donald
John A
JA
D000042:Absorption; D000818:Animals; D009320:Atrial Natriuretic Factor / Q000378:metabolism / Q000494:pharmacology; D001345:Autoradiography; D001667:Binding, Competitive; D001808:Blood Vessels / Q000502:physiology; D001834:Body Water / Q000378:metabolism; D002024:Bufo marinus; D002462:Cell Membrane / Q000378:metabolism; D003432:Cross-Linking Reagents; D006152:Cyclic GMP / Q000378:metabolism; D004848:Epithelium / Q000378:metabolism; D015870:Gene Expression; D007457:Iodine Radioisotopes; D009130:Muscle, Smooth / Q000378:metabolism; D020098:Natriuretic Peptide, C-Type / Q000378:metabolism / Q000494:pharmacology; D016133:Polymerase Chain Reaction; D011312:Pressure; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017461:Receptors, Atrial Natriuretic Factor / Q000235:genetics / Q000502:physiology*; D013552:Swine; D001743:Urinary Bladder / Q000098:blood supply / Q000378:metabolism*; D014668:Vasotocin / Q000494:pharmacology
D003432:Cross-Linking Reagents; D007457:Iodine Radioisotopes; D012333:RNA, Messenger; D020098:Natriuretic Peptide, C-Type; D009320:Atrial Natriuretic Factor; D017461:Receptors, Atrial Natriuretic Factor; D006152:Cyclic GMP; D014668:Vasotocin