Characteristics of ANP-binding sites in the adrenal capsules of term-pregnant rats

Abstract

Significant increases of circulatory volume and plasma aldosterone levels are observed in pregnancy. We investigated whether a decrease of atrial natriuretic peptide (ANP) receptors in the adrenal zona glomerulosa (ZG) could explain the marked elevation of plasma aldosterone occuring during pregnancy. 125I-ANP binding was measured in competition experiments using rANP(1–28), ANP(4–23), a truncated analog which has high specificity for the ANP-C receptor, or a combination of both. Western blot experiments were also performed with an investigation into the effect of ANP on aldosterone secretion in adrenal capsule suspensions. 125I-ANP binding on adrenal ZG membranes was displaced by ANP(1–28) with an affinity ( K d) of 313 ± 39 and 323 ± 60 pM (NS) for pregnant and non-pregnant rats, respectively. The density of sites ( B max) decreased slightly but not significantly during pregnancy. Displacement experiments with ANP(4–23) demonstrated a B max of 137 and 134 fmol/mg of proteins (NS) for pregnant and non pregnant rats, respectively. Studies were performed to block the ANP-C site. Displacing the remaining 125I-ANP binding with ANP(1–28) led to an affinity constant and receptor density which were not significantly different between the two groups of rats. The results obtained with Western blots showed a single band of 123 kDa with no significant variations in ANP-R 1 receptors in the ZG during gestation. The sensitivity of potassium-, ACTH- or angiotensin II-stimulated aldosterone secretion to ANP was not altered by gestation. These data show that the apparent hyperaldosteronism found in normal term-pregnant rats is not the consequence of modifications in the affinity, number and properties of ANP receptor types or in the sensitivity of the aldosterone response to ANP.