Potentiation of [Ca2+]i response to angiotensin III by cAMP in cortical thick ascending limb

Abstract

In the rat cortical thick ascending limb (CTAL), intracellular Ca2+ ([Ca2+]i) responses to angiotensin II (Ang II) and angiotensin III (Ang III) were mediated by the Ang II subtype 1A receptor (AT1A-R), whereas the arginine vasopressin (AVP)-dependent cAMP accumulation involved the vasopressin receptor type 2 (V2-R). This work was performed in CTAL to investigate the crosstalk between these two receptors by studying their transduction pathways. The cAMP-dependent pathway was activated by 10 minutes of prestimulation with either forskolin, CTP-cAMP or AVP, and Ang II/Ang III-induced [Ca2+]i responses were assessed. Pretreatment with 5 micromol/L forskolin significantly enhanced the [Ca2+]i response induced by 10-7 mol/L either Ang II or Ang III. Analysis of dose-response curves to Ang III in forskolin-treated CTAL demonstrated that the maximal [Ca2+]i response was significantly increased without altering the EC50. In Ca2+-free medium, the forskolin-induced potentiation of the [Ca2+]i response to Ang III was weaker but always present, suggesting that this effect was not only due to intracellular Ca2+ release but also to extracellular Ca2+ influx. Furthermore, the fact that the forskolin-induced potentiation of the [Ca2+]i response to Ang III was blocked by 10 micromol/L H-89, a specific protein kinase A (PKA) inhibitor, indicated that this effect occurred via activation of PKA. Finally, the potentiation of the [Ca2+]i response to Ang III also was observed following pretreatment with 100 micromol/L CTP-cAMP or 10-7 mol/L AVP. In CTAL, there is a positive crosstalk between the adenylyl cyclase and phosphoinositide pathways mediated by V2- and AT1A-R, respectively, through activation of PKA.