Mechanism of asynchronous Ca2+ waves underlying agonist‐induced contraction in the rat basilar artery

Background and purpose:  Uridine 5'-triphosphate (UTP) is a potent vasoconstrictor of cerebral arteries and induces Ca2+ waves in vascular smooth muscle cells (VSMCs). This study aimed to determine the mechanisms underlying UTP-induced Ca2+ waves in VSMCs of the rat basilar artery. Experimental approach:  Isometric force and intracellular Ca2+ ([Ca2+]i) were measured in endothelium-denuded rat basilar artery using wire myography and confocal microscopy respectively. Key results:  Uridine 5'-triphosphate (0.1–1000 µmol·L−1) concentration-dependently induced tonic contraction (pEC50 = 4.34 ± 0.13), associated with sustained repetitive oscillations in [Ca2+]i propagating along the length of the VSMCs as asynchronized Ca2+ waves. Inhibition of Ca2+ reuptake in sarcoplasmic reticulum (SR) by cyclopiazonic acid abolished the Ca2+ waves and resulted in a dramatic drop in tonic contraction. Nifedipine reduced the frequency of Ca2+ waves by 40% and tonic contraction by 52%, and the nifedipine-insensitive component was abolished by SKF-96365, an inhibitor of receptor- and store-operated channels, and KB-R7943, an inhibitor of reverse-mode Na+/Ca2+ exchange. Ongoing Ca2+ waves and tonic contraction were also abolished after blockade of inositol-1,4,5-triphosphate-sensitive receptors by 2-aminoethoxydiphenylborate, but not by high concentrations of ryanodine or tetracaine. However, depletion of ryanodine-sensitive SR Ca2+ stores prior to UTP stimulation prevented Ca2+ waves. Conclusions and implications:  Uridine 5'-triphosphate-induced Ca2+ waves may underlie tonic contraction and appear to be produced by repetitive cycles of regenerative Ca2+ release from the SR through inositol-1,4,5-triphosphate-sensitive receptors. Maintenance of Ca2+ waves requires SR Ca2+ reuptake from Ca2+ entry across the plasma membrane via L-type Ca2+ channels, receptor- and store-operated channels, and reverse-mode Na+/Ca2+ exchange. Mandarin translation of abstract