Noradrenaline-evoked cation conductance recorded with the nystatin whole-cell method in rabbit portal vein cells

Abstract

1. Noradrenaline-evoked currents were studied with the perforated patch technique (nystatin in the patch pipette) in freshly dispersed rabbit portal vein cells. 2. With potassium-containing solutions noradrenaline produced an inward current at -50 mV and an outward current (IK(Ca] at 0 mV. In potassium-free conditions noradrenaline elicited two distinct membrane currents which could be differentiated according to their sensitivity to the presence of caffeine in the bathing solution. One current was blocked whereas the second response was unaffected by caffeine. 3. The reversal potential (Er = -2.0 mV) of the caffeine-sensitive current was altered when external chloride was replaced with more permeant anions but Er was not changed when external sodium was replaced by Tris. Therefore the caffeine-sensitive current appears to be a calcium-activated chloride current (ICl(Ca]. 4. Er (+6.0 mV) of the caffeine-insensitive current was not altered by anion substitution but was changed when external sodium was replaced by Tris and barium ions. Thus the caffeine-insensitive conductance is a non-selective cation current (Icat). 5. When external NaCl was replaced by BaCl2, Er was shifted to more positive potentials which suggests that the cation conductance is more permeable to barium than to sodium. Icat was not affected by 10(-6) M-nifedipine. 6. The steady-state current-voltage relationship was linear between -50 and +50 mV for ICl(Ca) but the cation conductance mechanism displayed pronounced inward rectification and little outward current flowed across the membrane at positive potentials. 7. Caffeine (which releases calcium from internal stores) and the calcium ionophore ionomycin, which are expected to increase intracellular calcium concentration, evoked ICl(Ca) but not Icat. Thus ICl(Ca) but not Icat can be activated directly by an increase in intracellular calcium concentration. 8. When calcium was removed from the bathing solution the amplitude of Icat was not altered at early times in Ca(2+)-free conditions but was abolished after 10 min. Icat was readily activated by noradrenaline in the presence of ionomycin, which inactivated ICl(Ca). A permissive role for calcium in the generation of Icat is suggested. 9. The times between the application of noradrenaline and the onset of the chloride and cationic conductances were similar (0.75 and 0.9 s respectively). In contrast the rise time (3.9 s) and half-decay time (11 s) of Icat were much longer than the corresponding values of ICl(Ca) (respectively 1.9 and 2 s). In high-barium solution noradrenaline sometimes evoked a response that lasted for up to 10 min.(ABSTRACT TRUNCATED AT 400 WORDS)