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(Hypertension. 2007;49:618.)
© 2007 American Heart Association, Inc.

Original Articles, Part 2

Adenylyl Cyclase Isoform V Mediates Renin Release From Juxtaglomerular Cells

M. Cecilia Ortiz-Capisano; Pablo A. Ortiz; Pamela Harding; Jeffrey L. Garvin; William H. Beierwaltes

From the Hypertension and Vascular Research Division, Henry Ford Hospital, Detroit, Mich.

Correspondence to William H. Beierwaltes, Henry Ford Health System, 7121 E&R Bldg, 2799 W Grand Blvd, Detroit, MI 48202. E-mail wbeierw1{at}hfhs.org

We have shown previously that decreasing intracellular calcium in the juxtaglomerular cells increases both cAMP formation and renin release. We hypothesized that this is because of an interaction between intracellular calcium and the calcium-inhibitable isoform of adenylyl cyclase, type-V. We used primary cultures of juxtaglomerular cells isolated from C-57/B6 mice at 70% to 80% confluence. Western blots were performed on isolated juxtaglomerular cells using antibodies against either of the 2 calcium inhibitable isoforms of adenylyl cyclase, types-V and -VI. Only the antibody against adenylyl cyclase-V gave us a strong band at 120 kDa as expected. Immunolabeling in juxtaglomerular cells with confocal microscopy found immunofluorescence for the adenylyl cyclase-V–specific antibody compared with either negative controls or cells stained with the adenylyl cyclase-VI antibody. Reducing isolated juxtaglomerular intracellular calcium with 100 µmol/L of the cytosolic calcium chelator BAPTA-AM stimulated both cAMP (3.49±0.70 to 10.09±0.81 pmol/mL per milligram of protein; P<0.002) and renin release (1001.8±81.5 to 1648.0±139.1 ng of angiotensin I per milliliter per hour per milligram of protein; P<0.01). The selective adenylyl cyclase-V inhibitor NKY80 completely blocked both BAPTA-AM–stimulated cAMP formation and renin release. We conclude that lowering intracellular calcium is permissive, allowing an increased activity of the calcium-inhibitable isoform adenylyl cyclase-V (but not adenylyl cyclase-VI) in the juxtaglomerular cell, producing cAMP, which stimulates renin secretion.

Key Words: renin • adenylyl cyclase-V • calcium • juxtaglomerular cell

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