ATP-sensitive potassium channels in adult mouse skeletal muscle: different modes of blockage by internal cations, ATP and tolbutamide.

Single ATP-sensitive K channels were studied in membrane patches excised from enzymatically dissociated mouse toe muscle. The channel conductance is 74 pS in symmetrical 160 mM KCl solutions. Replacement of K+ by Na+ in the internal solution or 2 mM internal Ca2+ or Mg2+ induced a rectification of the current-voltage curve at positive potentials. No change of the current-voltage curve was observed by adding small amounts of the channel blockers ATP (20-100 microM) or tolbutamide (0.5 mM) to internal 160 mM KCl solutions. The openings of the channel occurred in bursts. Open (tau o), closed (tau c) times within bursts and pauses (tau p) between bursts were determined over a wide range of positive and negative membrane potentials. At increasing potentials tau o increases, tau c reaches a minimum near 0 mV and tau p decreases. According to the voltage dependence and the time scale of channel blockage three types of blocking agents could be distinguished: (i) small internal cations (Na+, Ca2+, Mg2+) are "fast" blockers at positive voltages; at negative voltages they decrease tau o and increase tau c. (ii) Internal ATP anions produce a voltage-dependent decline of the open-state probability and strongly decrease tau o. (iii) Tolbutamide causes a voltage-independent decrease of the open-probability and its main effect is an increase of tau p. The results suggest that the ATP-sensitive K channel has an internal gate like those of other voltage-gated cation channels and that different blockers interfere with different transitions in channel gating.