Enhancement of delayed rectifier K+ current by P2-purinoceptor stimulation in guinea-pig atrial cells

1. We studied the effects of P2-purinoceptor stimulation on the delayed rectifier K+ current (IK) in guinea-pig atrial myocytes using a whole-cell voltage-clamp technique. 2. External application of ATP increased IK, evoked by a 500 ms depolarizing pulse from a holding potential of -40 mV, under conditions in which the L-type Ca2+ channel was blocked; the effect was dose dependent with a half-maximal concentration (K1/2) of 0.95 microM. ATP (50 microM) produced a maximal increase of IK of about a factor of 2. 3. External ADP also enhanced IK in a dose-dependent manner with a K1/2 of 3.65 microM, whereas adenosine (100 microM) failed to evoke this response. Theophylline (500 microM), a blocker of the Pi-purinoceptor, did not antagonize the stimulating action of ATP on IK. These results indicate that IK was enhanced via P2-purinoceptors. 4. External ATP or ADP did not produce a significant change in the current kinetics of IK. 5. Pre-incubation of the atrial myocytes with pertussis toxin (PTX, 5 micrograms ml-1) did not affect the stimulating action of ATP on IK, indicating that PTX-sensitive G proteins did not mediate the ATP action. 6. The enhancement of IK by ATP developed slowly; the effects usually reached a maximum approximately 30-60 s after the application of ATP. This suggests the involvement of a diffusible cytosolic second messenger(s) in the response. ATP could further increase IK after maximal enhancement by isoprenaline (0.5-1.0 microM), suggesting that the intermediate steps were independent of cyclic AMP-dependent protein kinase (protein kinase A). 7. Potentiation of IK by ATP was not attenuated by either (i) pretreatment of the cells with 5 microM 1-(5-isoquinolinylsulphonyl)-2-methylpiperazine dihydrochloride (H-7) or (ii) intracellular perfusion of 20 mM 1,2-bis(O-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA), suggesting that protein kinase C and intracellular Ca2+ did not mediate the response. 8. It is concluded that the activation of P2-purinoceptors increases IK through intracellular mechanisms independent of protein kinase A, protein kinase C or intracellular free Ca2+ in guinea-pig atrial myocytes.