Kinetics of ATP-induced Ca2+ transients in cultured pig aortic smooth muscle cells depend on ATP concentration and stored Ca2+.

1. Single cultured pig aortic smooth muscle cells were studied using fura-2 and dual excitation wavelength microfluometry. 2. Extracellular ATP in micromolar concentrations induced a transient increase of [Ca2+]i due to Ca2+ release from internal stores. In the same concentration range application of ATP resulted in an increase of intracellular inositol phosphate level. 3. In a medium range of ATP concentrations (2-10 microM) the Ca2+ signal was oscillating, whereas at higher and lower concentrations only a Ca2+ transient with a single peak was elicited. 4. The rank order of potency for the tested purine and pyrimidine nucleotides was: UTP > ATP > ADP >> AMP = adenosine = alpha,beta-methylene ATP = 0. The response to the nucleotides could be abolished by the P2-purinoceptor antagonist suramin. 5. The latency between agonist application and onset of the Ca2+ transients as well as their amplitude and rate of rise are dependent on ATP concentration. 6. Removal of Ca2+ from the extracellular solution led to a progressive decrease of amplitude and prolonged latency of the Ca2+ transients. This shows that depletion of the Ca2+ stores affects kinetics of the ATP-induced Ca2+ release. 7. The inorganic Ca(2+)-influx blockers Ni2+ and Co2+ affected amplitude and latency in a manner similar to Ca2+ removal, while the Ca2+ antagonist nifedipine was ineffective up to a concentration of 10(-6) M. 8. These results reveal a dual dependency of the InsP3-induced Ca2+ release on agonist concentration and filling state of the Ca2+ stores, which supports the hypothesis of a feedback amplification between InsP3 and released Ca2+.