Openers of small conductance calcium-activated potassium channels selectively enhance NO-mediated bradykinin vasodilatation in porcine retinal arterioles

Background and purpose:

Small (SK_Ca or K_Ca2) and intermediate (IK_Ca or K_Ca3.1) conductance calcium-activated potassium channels are involved in regulation of vascular tone and blood pressure. The present study investigated whether NS309 (6,7-dichloro-1H-indole-2,3-dione 3-oxime) and CyPPA (cyclohexyl-[2-(3,5-dimethyl-pyrazol-1-yl)-6-methyl-pyrimidin-4-yl]-amine), which are selective openers of SK_Ca and IK_Ca channels and of SK_Ca2 and SK_Ca3 channels, respectively, enhance endothelium-dependent vasodilatation in porcine retinal arterioles.

Experimental approach:

In porcine retinal arterioles, SK_Ca3 and IK_Ca protein localization was examined by immunolabelling. Endothelial cell calcium was measured by fluorescence imaging. For functional studies, arterioles with internal diameters of 116 ± 2 µm (n = 276) were mounted in microvascular myographs for isometric tension recordings.

Key results:

SK_Ca3 and IK_Ca protein was localized in the endothelium. Bradykinin, but not NS309 or CyPPA increased endothelial cell calcium. Pre-incubation with NS309 or CyPPA enhanced bradykinin relaxation without changing endothelial cell calcium. This enhanced relaxation was abolished by blocking SK_Ca channels with apamin. In the presence of NS309 or CyPPA, mainly inhibition of NO synthase with asymmetric dimethylarginine, but also inhibition of cyclooxygenase with indomethacin, reduced bradykinin relaxation. Bradykinin relaxation was completely abolished by NO synthase and cyclooxygenase inhibition together with a NO scavenger, oxyhaemoglobin.

Conclusions and implications:

In porcine retinal arterioles, bradykinin increases endothelial cell calcium leading to activation of SK_Ca and IK_Ca channels. Without altering endothelial cell calcium, NS309 and CyPPA open SK_Ca channels that enhance NO-mediated bradykinin relaxations. These results imply that opening SK_Ca channels improves endothelium-dependent relaxation and makes this channel a potential target for treatments aimed at restoring retinal blood flow.