ATP-activated cation currents in single guinea-pig hepatocytes

1. Responses of single guinea-pig liver cells to the application of external ATP were studied using the whole-cell voltage clamp technique. 2. When the cells were loaded with 5 mM EGTA in the absence of K+ and Cl- in both internal and external solutions, application of ATP (0.03-100 mu M) elicited a. large cation-selective inward current at negative holding potentials. The current densities at the peak of the response to 100 mu M ATP were 4.5 +/- 0.5 pA pF(-1) (mean +/- S.E.M., n = 18) in the presence of Na+ and Ca2+ in the external medium and 3.3 +/- 0.7 pA pF(-1) (n = 6) with Ca2+ as the major permeant ion. 3. Divalent cations, when added during the response to ATP in the presence of Na+ and Ca2+, exerted different effects: CdSO4 (2 mM) totally and NiSO4 (2 mM) partially blocked the inward current whereas MnSO4 (2 mM) did not block it. The ATP-activated conductance was permeable to all the divalent cations tested in this study, i.e. Ca2+, Cd2+, Ni2+, Mn2+ and Mg2+. No response to ATP was observed in the absence of external cations. 4. The activation of the inward current was not maintained in the continuous presence of ATP. The effect of Ca2+ ions on the desensitization of the response was studied in different external solutions. The decline in the amplitude of the inward current after the peak was fitted with a single exponential with a time constant of about 2 s for pure Ca2+, Cd2+ or Ni2+ currents, 3 s for Mg2+ or Mn2+ and 4 s in the presence of both Na+ and Ca2+. 5. Under more physiological conditions, the entry of Ca2+ evoked after the stimulation of P2X purinoceptors was associated with an increase in fluo-3 fluorescence and a marked reduction in the delay before the mobilization of internal Ca2+ stores following the activation of P2Y purinoceptors.