ATP regulates muscarine-sensitive potassium current in dissociated bull-frog primary afferent neurones

1. Bull-frog dorsal root ganglion cells in primary culture were voltage clamped in the whole-cell configuration. The pipette solution contained ATP (5 mM). 2. Step depolarizations (5-70 mV, 0.1-1 s) from a holding potential close to the resting potential (range, -64 to -79 mV) evoked a non-inactivating potassium current with properties indistinguishable from those which have been reported for the M-current of bull-frog sympathetic neurones. 3. An unhydrolysable ATP analogue APP(NH)P (5 mM), substitute with ATP in the pipette solution, did not support the M-current activation. 4. Bath application of ATP (30 nM-30 microM) reduced the amplitude of the M-current in a concentration-dependent manner, congruent to 50% inhibition of the current occurring with 1 microM-ATP. The main effect of ATP was to reduce the maximum M-conductance without changing the activation and deactivation kinetics of the M-current. 5. Essentially the same results were obtained with ADP (0.1-30 microM) and alpha, beta-methylene-ATP (10-30 microM). AMP (10-100 microM) and adenosine (10-30 microM) were without effect on the M-current. 6. The ATP-induced inhibition of the M-current was irreversible when an unhydrolysable GTP analogue GTP-gamma-S (10-30 microM) was present in the pipette solution. ATP (3 microM) reduced the amplitude of the M-current only by about 10% when GDP-beta-S (100 microM) was present in the pipette solution. Pre-treatment of the cells with pertussis toxin (IAP; 500 ng ml-1) for 24 h at 24 degrees C did not prevent the ATP-induced M-current inhibition. 7. Phorbol 12-myristate 13-acetate (PMA; 1-3 microM) reduced the amplitude of the M-current to about 50%. A reduction in the M-current amplitude by PMA (3 microM) and ATP (10 microM) was attenuated when staurosporine (200 nM) was present in the pipette solution. Forskolin (10 microM) was without effect on the M-current. 8. It is concluded that ATP acting at P2 receptors, associated with an IAP-insensitive GTP-binding protein, inhibits the M-current in amphibian primary afferent neurones.