Molecular mechanism of ADP-ribosyl cyclase activation in angiotensin II signaling in murine mesangial cells

ADP-ribosyl cyclase (ADPR-cyclase) produces a [Ca.sup.2+]-mobilizing second messenger cyclic ADP-ribose (cADPR) from NA[D.sup.+]. In this study, we investigated the molecular basis of ADPR-cyclase activation and the following cellular events in angiotensin II (ANG II) signaling in mouse mesangial cells (MMCs). Treatment of MMCs with ANG II induced an increase in intracellular [Ca.sup.2+] concentrations through a transient [Ca.sup.2+] release via an inositol 1,4,5-trisphosphate receptor and a sustained [Ca.sup.2+] influx via L-type [Ca.sup.2+] channels. The sustained [Ca.sup.2+] signal, but not the transient [Ca.sup.2+] signal, was blocked by a cADPR antagonistic analog, 8-bromo-cADPR (8-Br-cADPR), and an ADPRcyclase inhibitor, 4,4'-dihydroxyazobenzene (DHAB). In support of the results, ANG II stimulated cADPR production in a time-dependent manner, and DHAB inhibited ANG II-induced cADPR production. Application of pharmacological inhibitors revealed that activation of ADPR-cyclase by ANG II involved ANG II type 1 receptor, phosphoinositide 3-kinase, protein tyrosine kinase, and phospolipase C-[gamma]1. Moreover, DHAB as well as 8-Br-cADPR abrogated ANG II-mediated Akt phosphorylation, nuclear translocation of nuclear factor of activated T cell, and uptake of [[sup.3]H]thymidine and [[sup.3]H]leucine in MMCs. These results demonstrate that ADPR-cyclase in MMCs plays a pivotal role in ANG II signaling for cell proliferation and protein synthesis. cyclic ADP-ribose; intracellular [Ca.sup.2+] concentration