DiBAC4(3) hits a “sweet spot” for the activation of arterial large-conductance Ca2+-activated potassium channels independently of the β1-subunit

Abstract

The voltage-sensitive dye bis-(1,3-dibutylbarbituric acid)trimethine oxonol [DiBAC₄(3)] has been reported as a novel large-conductance Ca²⁺-activated K⁺ (BK) channel activator with selectivity for its β₁- or β₄-subunits. In arterial smooth muscle, BK channels are formed by a pore-forming α-subunit and a smooth muscle-abundant regulatory β₁-subunit. This tissue specificity has driven extensive pharmacological research aimed at regulating arterial tone. Using animals with a disruption of the gene for the β₁-subunit, we explored the effects of DiBAC₄(3) in native channels from arterial smooth muscle. We tested the hypothesis that, in native BK channels, activation by DiBAC₄(3) relies mostly on its α-subunit. We studied BK channels from wild-type and transgenic β₁-knockout mice in excised patches. BK channels from brain arteries, with or without the β₁-subunit, were similarly activated by DiBAC₄(3). In addition, we found that saturating concentrations of DiBAC₄(3) (~30 μM) promote an unprecedented persistent activation of the channel that negatively shifts its voltage dependence by as much as -300 mV. This "sweet spot" for persistent activation is independent of Ca²⁺ and/or the β₁₋₄-subunits and is fully achieved when DiBAC₄(3) is applied to the intracellular side of the channel. Arterial BK channel response to DiBAC₄(3) varies across species and/or vascular beds. DiBAC₄(3) unique effects can reveal details of BK channel gating mechanisms and help in the rational design of BK channel activators.