Whole-cell and single-channel currents across the plasmalemma of corn shoot suspension cells

Abstract

Whole-cell sealed-on pipettes have been used to measure electrical properties of the plasmalemma surrounding protoplasts isolated from Black Mexican sweet corn shoot cells from suspension culture. In these protoplasts the membrane resting potential (Vm) was found to be -59 +/- 23 mV (n = 23) in 1 mM Ko+. The mean Vm became more negative as [K+]o decreased, but was more positive than the K+ equilibrium potential. There was no evidence of electrogenic pump activity. We describe four features of the current-voltage characteristic of the plasmalemma of these protoplasts which show voltage-gated channel activity. Depolarization of the whole-cell membrane from the resting potential activates time- and voltage-dependent outward current through K(+)-selective channels. A local minimum in the outward current-voltage curve near Vm = 150 mV suggests that these currents are mediated by two populations of K(+)-selective channels. The absence of this minimum in the presence of verapamil suggests that the activation of one channel population depends on the influx of Ca2+ into the cytoplasm. We identify unitary currents from two K(+)-selective channel populations (40 and 125 pS) which open when the membrane is depolarized; it is possible that these mediate the outward whole-cell current. Hyperpolarization of the membrane from the resting potential produces time- and voltage-dependent inward whole-cell current. Current activation is fast and follows an exponential time course. The current saturates and in some cases decreases at membrane potentials more negative than -175 mV. This current is conducted by poorly selective K+ channels, where PCl/PK = 0.43 +/- 0.15. We describe a low conductance (20 pS) channel population of unknown selectivity which opens when the membrane is hyperpolarized. It is possible that these channels mediate inward whole-cell current. When the membrane is hyperpolarized to potentials more negative than -250 mV large, irregular inward current is activated. A third type of inward whole-cell current is briefly described. This activates slowly and with a U-shaped current-voltage curve over the range of membrane potentials -90 less than Vm less than 0 mV.