Calcium oscillations induced by ATP in human umbilical cord smooth muscle cells

Arterial smooth muscle cells exhibit vasomotion, related to oscillations in intracellular Ca(2+) concentration, but the origin and function of these has not yet been fully determined. We measured intracellular Ca(2+) using conventional fluorescent methods in primary cultured, human umbilical cord artery smooth muscle cells (HUCASMC). Spontaneous oscillations in Ca(2+) was found in only 1% of all cells but exogenous, micromolar concentrations of ATP could induce Ca(2+) oscillations in 70% of cells with the most common pattern being one of regular amplitude and frequency with a return to basal levels between each peak. The P2Y agonist, UTP, but not the P2X agonist alphabeta-methylene ATP, could also induce Ca(2+) oscillations. Once induced, these oscillations could not be blocked by G-protein, PLC, VGCC or TRP channel antagonists applied individually, but could be prevented when antagonists were applied together. In the presence of EGTA, micromolar concentrations of ATP induced an elevation in intracellular Ca(2+) but did not induce Ca(2+) oscillations. The oscillation frequency induced by ATP was affected by bath Ca(2+) concentration. Taken together, these data suggest that external Ca(2+) entry maintains the Ca(2+) oscillation induced by activation of P2Y receptors. Once induced, multiple mechanisms are involved to maintain the oscillation and the oscillation frequency is determined by the speed of Ca(2+) refilling. Chronic hypoxia enhanced the Ca(2+) response and altered the oscillation frequency. We suggest that these oscillations may play a role in the maintenance of umbilical blood flow during situations in which GPCR are activated.