Detection of a receptor for angiotensinogen on placental cells

• Published in: American journal of hypertension 2003-01, Vol.16 (1), p.59-62
• Main Authors: Tewksbury, Duane A, Pan, Nan, Kaiser, Steven J
• Format: Article
• Language: English
• Subjects: Angiotensin; Angiotensinogen - metabolism; Angiotensinogen - pharmacology; Arterial hypertension. Arterial hypotension; Binding, Competitive; Biological and medical sciences; Blood and lymphatic vessels; Cardiology. Vascular system; Cell Line; Clinical manifestations. Epidemiology. Investigative techniques. Etiology; Humans; Iodine Radioisotopes; Medical sciences; Placenta - cytology; Placenta - metabolism; Radioligand Assay; receptor; Receptors, Cell Surface - metabolism; renin-angiotensin system; Renin-Angiotensin System - physiology
• ISSN: 0895-7061; 1879-1905; 1941-7225
• DOI: 10.1016/S0895-7061(02)03079-0

Current evidence indicates that there may be a tissue-specific renin-angiotensin system (RAS) in the human placenta. To better define the placental RAS, this study sought to determine whether placental derived cells possessed a receptor for angiotensinogen (AGT), a rate limiting component of the RAS. A human placenta–derived cell line, CRL-7548, a highly purified AGT and iodine-125–labeled angiotensinogen (125I-AGT) were used in this study. The cells were seeded in 35-mm diameter plastic wells, cultured for 2 days in fetal bovine serum–Dulbecco’s modified Eagle’s medium to reach about 80% to 90% confluence, 2 × 104 cells/well and passed in Dulbecco’s modified Eagle’s medium free of fetal bovine serum before experimentation. A quantity of 3 μg of 125I-AGT was added to each well at zero time. The cells rapidly bound 125I-AGT in a time dependent manner with saturation being achieved in 2 to 4 h. This binding was competitively inhibited by unlabeled AGT. Prior addition of a 100-fold excess of unlabeled AGT resulted in a 54% decrease in maximal binding. Bound AGT was also competitively displaced by AGT. Addition of a 200-fold excess of unlabeled AGT displaced 70% of the bound 125I-AGT. Thus, approximately 30% of the total binding can be attributed to nonspecific binding. An acid wash (0.2 mol/L acetic acid, 0.5 mol/L NaCl, pH 2.5) at 4°C removed 46% of the bound 125I-AGT. An acid wash is known to dissociate cell surface ligand-receptor complexes, but does not remove internalized ligand. The results of this study provide evidence for the existence of an AGT receptor on placenta-derived cells. This is the first report of an AGT receptor.