A novel angiotensin receptor subtype in rat mesangium. Coupling to adenylyl cyclase.
The diversity of angiotensin II (Ang II) actions implies multiple receptor subtypes. To characterize these subtypes in rat mesangial cells, we used the angiotensin subtype 1A (AT1A) antagonist losartan (DuP 753), the subtype 2/1B (AT2/AT1B) antagonist PD 123319, and the AT2 antagonist CGP 42112A in radioreceptor and adenylyl cyclase assays. In radioligand binding competition experiments, approximately 25% of the specific binding sites labeled by 125I-[Sar1]Ang II were inhibited by low concentrations of PD 123319 (0.1 to 10 nM), whereas the AT2 antagonist CGP 42112A was inactive at concentrations less than 0.1 microM. Conversely, losartan inhibited 75% of the binding at low concentrations (0.1 nM to 0.1 microM), but higher concentrations (up to 10 microM) were required to inhibit the second component of 125I-[Sar1]Ang II binding. The effects of the different antagonists on the inhibition by Ang II of forskolin-stimulated cyclic AMP production were also analyzed. Ang II inhibited forskolin-stimulated adenylyl cyclase in a concentration-dependent fashion (IC50, 35 +/- 7 nM), and the maximal inhibition of adenylyl cyclase was 44 +/- 2%. In the radioligand binding experiments, both losartan and PD 123319 antagonized the inhibition of adenylyl cyclase elicited by 0.1 microM Ang II (IC50, 0.5 +/- 0.2 and 1.2 +/- 0.4 microM, respectively), whereas CGP 42112A was less potent (IC50, 5.7 +/- 1.6 microM). Comparison of binding affinities at AT1B receptor sites with antagonist potencies in the adenylyl cyclase assay show good agreement for losartan and CGP 42112A, whereas PD 123319 is less potent than expected from membrane binding assays, possibly because of partial agonist properties.(ABSTRACT TRUNCATED AT 250 WORDS)
Department of Medicine, School of Medicine, Case Western Reserve University, Cleveland, Ohio 44106-4982.
J
J
Zhou
P
P
Ernsberger
J G
JG
Douglas
D000067956:Adenylyl Cyclase Inhibitors; D000262:Adenylyl Cyclases / Q000378:metabolism*; D000804:Angiotensin II / Q000494:pharmacology; D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism; D005920:Glomerular Mesangium / Q000166:cytology / Q000378:metabolism*; D051381:Rats; D011945:Receptors, Angiotensin / Q000378:metabolism*
D000067956:Adenylyl Cyclase Inhibitors; D011945:Receptors, Angiotensin; D000804:Angiotensin II; D005576:Colforsin; D000242:Cyclic AMP; D000262:Adenylyl Cyclases