Bidirectional TRP/L Type Ca²⁺ Channel/RyR/BK_Ca Molecular and Functional Signaloplex in Vascular Smooth Muscles

TRP channels are expressed both in vascular myocytes and endothelial cells, but knowledge of their operational mechanisms in vascular tissue is particularly limited. Here, we show for the first time the biphasic contractile reaction with relaxation followed by a contraction in response to TRPV4 agonist, GSK1016790A, in a rat pulmonary artery preconstricted with phenylephrine. Similar responses were observed both with and without endothelium, and these were abolished by the TRPV4 selective blocker, HC067047, confirming the specific role of TRPV4 in vascular myocytes. Using selective blockers of BK_Ca and L-type voltage-gated Ca²⁺ channels (Ca_L), we found that the relaxation phase was inducted by BK_Ca activation generating STOCs, while subsequent slowly developing TRPV4-mediated depolarisation activated Ca_L, producing the second contraction phase. These results are compared to TRPM8 activation using menthol in rat tail artery. Activation of both types of TRP channels produces highly similar changes in membrane potential, namely slow depolarisation with concurrent brief hyperpolarisations due to STOCs. We thus propose a general concept of bidirectional TRP-Ca_L-RyR-BK_Ca molecular and functional signaloplex in vascular smooth muscles. Accordingly, both TRPV4 and TRPM8 channels enhance local Ca²⁺ signals producing STOCs via TRP-RyR-BK_Ca coupling while simultaneously globally engaging BK_Ca and Ca_L channels by altering membrane potential.