Current-voltage relationships and voltage sensitivity of the Cl⁻ pump inHalicystis parvula

Summary

The current-voltage (I–V) relationships for internally perfused and nonperfused cells ofHalicystis parvula were determined. In both types of cells theI–V curve shows a conspicuous region of negative slope, beginning at vacuole potentials around −30 mV and continuing to values of +20 to +40mV. The negative slope in perfused cells is abolished by the metabolic inhibitors, darkness and low temperature. In order to determine the origin of this negative slope, we measured the voltage sensitivity of the unidirectional fluxes of Cl⁻, Na⁺ and K⁺ in perfused cells. The results show that the Cl⁻ influx, which is mediated primarily by a Cl⁻ pump, increases as the vacuole potential is clamped at increasingly morenegative values up to −50 mV, while the other fluxes measured changed in the directions predicted by the change in electrical driving force. The voltage sensitivity of the Cl⁻ pump quantitatively accounts for the negative slope of theI–V curve. Also, we observed a large transient outward current of 10–20-sec duration following an abrupt depolarization by voltage clamping. This transient current was reduced or abolished by low temperature, which suggests that it may be due to the voltage-sensitive Cl⁻ pump. Finally, we found an inverse relationship between the transprotoplasm resistance (R _m ) and thePD under standard conditions, which suggests that the activity of the electrogenic Cl⁻ pump lowerR _m , i.e., it is a conductive pump.