The Adenylyl Cyclase Inhibitor MDL-12,330A Potentiates Insulin Secretion via Blockade of Voltage-Dependent K(+) Channels in Pancreatic Beta Cells

OBJECTIVE: Adenylyl cyclases (ACs) play important role in regulating pancreatic beta cell growth, survival and secretion through the synthesis of cyclic AMP (cAMP). MDL-12,330A and SQ 22536 are two AC inhibitors used widely to establish the role of ACs. The goal of this study was to examine the effects of MDL-12,330A and SQ 22536 on insulin secretion and underlying mechanisms. METHODS: Patch-clamp recording, Ca(2+) fluorescence imaging and radioimmunoassay were used to measure outward K(+) currents, action potentials (APs), intracellular Ca(2+) ([Ca(2+)](i)) and insulin secretion from rat pancreatic beta cells. RESULTS: MDL-12,330A (10 µmol/l) potentiated insulin secretion to 1.7 times of control in the presence of 8.3 mmol/l glucose, while SQ 22536 did not show significant effect on insulin secretion. MDL-12,330A prolonged AP durations (APDs) by inhibiting voltage-dependent K(+) (K(V)) channels, leading to an increase in [Ca(2+)](i) levels. It appeared that these effects induced by MDL-12,330A did not result from AC inhibition, since SQ 22536 did not show such effects. Furthermore, inhibition of the downstream effectors of AC/cAMP signaling by PKA inhibitor H89 and Epac inhibitor ESI-09, did not affect K(V) channels and insulin secretion. CONCLUSION: The putative AC inhibitor MDL-12,330A enhances [Ca(2+)](i) and insulin secretion via inhibition of K(V) channels rather than AC antagonism in beta cells, suggesting that the non-specific effects is needed to be considered for the right interpretation of the experimental results using this agent in the analyses of the role of AC in cell function.