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Ca 2+ -dependent potassium channels and cannabinoid signaling in the endothelium of apolipoprotein E knockout mice before plaque formation.
Journal of Molecular and Cellular Cardiology 2018 Februrary
Endothelial Ca2+ -dependent K+ channels (KCa ) regulate endothelial function. We also know that stimulation of type 2 cannabinoid (CB2 ) receptors ameliorates atherosclerosis. However, whether atherosclerosis is accompanied by altered endothelial KCa - and CB2 receptor-dependent signaling is unknown. By utilizing an in situ patch-clamp approach, we directly evaluated the KCa channel function and the CB2 receptor-dependent electrical responses in the endothelium of aortic strips from young ApoE-/- and C57Bl/6 mice. In the ApoE-/- group, the resting membrane potential (-30.1±1.1mV) was less negative (p<0.05) compared to WT (-38.9±1.4mV) and voltage ramps generated an overall KCa current of reduced amplitude. The peak hyperpolarization to 2μM Ach was not different between the groups. However, the sustained component was significantly reduced in ApoE-/- strips. In contrast, the peak hyperpolarization to 0.2μM Ach was increased in the ApoE-/- group, and SKA-31, a direct IKCa /SKCa channel opener, produced a hyperpolarization and whole-cell current of greater amplitude. The BKCa opener NS1619 produced hyperpolarization that was enhanced in ApoE-/- group. N-arachidonoyl glycine, a BKCa opener, produced a hyperpolarization of enhanced amplitude in ApoE-/- arteries. Selective CB2 receptor agonist AM1241 (5μM) had no effect on endothelial membrane potential in WT group; however, in ApoE-/- group, it elicited hyperpolarization that was inhibited by a selective CB2 receptor antagonist AM630. Conclusively, our data point to functional down-regulation of basal IKCa activity in unstimulated endothelium of ApoE-/- mice. Direct and indirect IKCa stimulation resulted in increased recruitment of the channels. In addition, our data point to up-regulation of endothelial BKCa channels and CB2 receptors in ApoE-/- arteries.
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