Extracellular ATP directly gates a cation-selective channel in rabbit airway ciliated epithelial cells.

Abstract

1. A membrane conductance activated by extracellular ATP was identified and characterized in freshly dissociated rabbit airway ciliated cells using the whole-cell and outside-out patch configurations of the patch-clamp technique. 2. In solutions designed to maximize currents through voltage-gated calcium channels, there were no indications of voltage-gated Ba2+ currents. 3. Extracellular ATP (but not UTP or ADP) activated a membrane conductance which remained activated for several minutes in the presence of ATP. The conductance was permeable to monovalent and divalent cations with approximate relative permeabilities (P) for PBa : PCs : PTEA of 4 : 1 : 0.1. Permeability to Cl- was negligible. 4. Including GDP-beta-S in the intracellular solution did not inhibit the effects of ATP, nor did GTP-gamma-S irreversibly activate the conductance. 5. In outside-out membrane patches, with GDP-beta-S in the pipette solution, ATP activated ion channels which had a chord conductance of approximately 6 pS in symmetrical 150 mM CsCl solutions at -120 mV. 6. Suramin (100 microM) inhibited the whole-cell currents activated by ATP (200 microM) by 93 +/- 3 %. Similar effects of suramin were observed on ATP-activated channels in outside-out membrane patches. 7. Extracellular ATP had a priming action on the response to subsequent exposure to ATP. At -40 mV, the time to half-maximal current activation (t1/2) was 46 +/- 9 s during the first exposure to 200 microM ATP and decreased to 5 +/- 3 s during a second exposure to the same concentration of ATP. The priming action of ATP was not inhibited by including GDP-beta-S in the intracellular solution. 8. The initial rate of activation increased with the concentration of ATP, and was voltage sensitive. During the first exposure to 200 microM ATP, t1/2 at +40 mV was 4-fold longer than t1/2 at -40 mV. 9. Half-maximal activation of the conductance shifted from 210 +/- 30 to 14 +/- 4 microM added ATP when CaCl2 in the extracellular solution was reduced from 1.58 to 0. 01 mM. The Hill coefficient for ATP was 1.2 in both solutions.10. These observations suggest that a form of ATP uncomplexed with divalent cations directly gates an ion channel (P2X receptor) in rabbit airway ciliated cells, which serves as a pathway for Ca2+ influx. This purinoceptor may contribute to sustained ciliary activation during prolonged exposures to ATP.