Sodium--potassium Pump Current in Rabbit Sino-atrial Node Cells

1. The Na(+)-K+ pump current (Ip) was studied in sino-atrial (SA) node cells of rabbits using the whole-cell patch-clamp technique. 2. With 50 mM Na+ in the pipette solution ([Na+]pip), changing the external K+ concentration (-K+-o) from 0 to 5.4 mM caused the holding current to shift in an outward direction and reach a new steady state. The current-voltage relationships obtained by subtraction of current traces recorded at 0 mM Ko+ from those recorded at 5.4 mM Ko+ revealed time-independent and voltage-dependent characteristics. The external K(+)-induced current was completely blocked by external application of 10 microM ouabain, indicating the existence of Ip in SA node cells of rabbit heart. 3. Ip increased as [K+]o increased. With 30 mM Na+pip, Ip at 0 mV was activated by [K+]o with non-linear least-squares fit parameters for the Hill equation of K0.5 of 1.4 mM and a Hill coefficient (nH) of 1.2 (n = 7). 4. The cation dependence of the K+ site of the Na(+)-K+ pump was examined using various monovalent cations. The sequence was K+ > or = Rb+ > Cs+ > > > Li+. 5. Ip at 0 mV also increased as [Na+]pip was increased from 10 to 150 mM at 5.4 mM Ko+, with a K0.5 value of 14 mM and a nH of 1.3 (n = 54). 6. Ip at 0 mV was reduced by lowering the temperature from 37 to 25 degrees C with 30 mM Na+pip and 5.4 mM Ko+. The temperature coefficient (Q10) for Ip was 2.1 (n = 27). 7. With 10 mM Na+pip and 5.4 mM Ko+, the half-activation voltage of Ip was -52 +/- 16 mV and the current at this voltage was 22.5 +/- 3.5 pA (n = 10), indicating that Ip contributes significantly to the background outward current during the normal pacemaker potential of SA node cells.