Distribution and characterization of natriuretic peptide receptors in the kidney of the toad, Bufo marinus.
The location and characteristics of atrial natriuretic peptide binding sites in the kidney of the toad, Bufo marinus, were determined. Specific (125)I-rANP binding sites were observed on glomeruli and blood vessels, but little if any binding was observed over regions corresponding to the renal tubules. (125)I-rANP binding in tissue sections and/or isolated membranes was completely displaced in the presence of 1 microM rat ANP, frog ANP, and porcine C-type natriuretic peptide (membranes only); however, residual binding remained after incubation with 1 microM of the NPR-C ligand, C-ANF, indicating the presence of two distinct binding sites. Electrophoresis of kidney membranes cross-linked to (125)I-rANP identified specific bands at approximately 70 and 140 kDa which correspond to the monomeric mass of NPR-C and the guanylate cyclase receptors, respectively. In addition, rat ANP, frog ANP, and porcine CNP stimulated a significant increase in cGMP production rates in membrane preparations, while C-ANF had no stimulatory effect. Two partial cDNA clones generated using primers based on conserved regions of vertebrate natriuretic peptide receptors showed high homology to an NPR-C and the natriuretic peptide guanylate cyclase receptors (NPR-GC), respectively. This study provides evidence that the kidney of B. marinus contains both NPR-C and NPR-GC and that the glomerulus is potentially the principal site of ANP regulation in the kidneys.
School of Biological and Chemical Sciences, Deakin University, Geelong, Victoria, 3217, Australia.
S K
SK
Meier
T
T
Toop
J A
JA
Donald
D000595:Amino Acid Sequence; D000818:Animals; D001345:Autoradiography; D001483:Base Sequence; D001667:Binding, Competitive / Q000187:drug effects; D002024:Bufo marinus / Q000502:physiology*; D003001:Cloning, Molecular; D003432:Cross-Linking Reagents; D004591:Electrophoresis, Polyacrylamide Gel; D006162:Guanylate Cyclase / Q000378:metabolism; D007668:Kidney / Q000033:anatomy & histology / Q000201:enzymology / Q000378:metabolism*; D008024:Ligands; D008566:Membranes / Q000737:chemistry / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D017461:Receptors, Atrial Natriuretic Factor / Q000378:metabolism*
D003432:Cross-Linking Reagents; D008024:Ligands; D012333:RNA, Messenger; D006162:Guanylate Cyclase; D017461:Receptors, Atrial Natriuretic Factor