Regulation of atrial natriuretic peptide receptors in vascular smooth muscle cells: role of cGMP.
We tested the hypothesis that intracellular guanosine 3',5'-cyclic monophosphate (cGMP) regulates atrial natriuretic peptide (ANP) receptors. The effect of chronic exposure to ANP, sodium nitroprusside (SNP), and 8-bromo-cGMP (8-BrcGMP) on ANP receptors and cGMP formation was determined in guinea pig thoracic aorta smooth muscle cells (TASM) and in coronary artery smooth muscle cells (CASM). TASM express both the ANP-activated guanylyl cyclase (B-receptor) and the clearance receptor (C-receptor) and respond to ANP with increased cGMP. CASM exhibit only the ANP C-receptor. In TASM, 24-h treatment with 1 microM atriopeptin (AP) III [rat ANP-(103-126)] caused a sixfold increase in basal cGMP levels, which were unaltered in CASM. In AP III-treated TASM, maximal binding of 125I-labeled AP III (Bmax) was reduced 40% while affinity [dissociation constant (KD)] was unaltered. In CASM, Bmax and KD were not affected. In treated TASM, washed free of AP III, acute dose-response curves of cGMP to AP III were not different from that in untreated cells. In both TASM and CASM, basal cGMP levels were elevated and Bmax was decreased by 6-h treatment with SNP. SNP did not alter the acute response of cGMP to AP III. In both cell types, 8-BrcGMP for 6 h caused reduction in Bmax. These findings support the conclusion that elevated cGMP was necessary for ANP-induced downregulation of receptors. The unaltered responsiveness of cGMP to AP III suggests the B-receptor was not altered and the reduced Bmax was due to decreased C-receptor.
Medical Center of Central Georgia, Mercer University, Macon 31201.
L M
LM
Zhang
H
H
Tao
W H
WH
Newman
D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology; D000818:Animals; D009320:Atrial Natriuretic Factor / Q000378:metabolism / Q000494:pharmacology; D002478:Cells, Cultured; D006152:Cyclic GMP / Q000502:physiology*; D004305:Dose-Response Relationship, Drug; D007425:Intracellular Membranes / Q000378:metabolism; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism*; D009599:Nitroprusside / Q000494:pharmacology; D009994:Osmolar Concentration; D010446:Peptide Fragments; D017461:Receptors, Atrial Natriuretic Factor / Q000378:metabolism*; D013997:Time Factors
D010446:Peptide Fragments; D009599:Nitroprusside; D015124:8-Bromo Cyclic Adenosine Monophosphate; C062525:atrial natriuretic factor prohormone (103-126); D009320:Atrial Natriuretic Factor; D017461:Receptors, Atrial Natriuretic Factor; D006152:Cyclic GMP