Abstract
Two voltage-dependent inward currents in the fresh-water hypotrichous ciliate Stylonychia mytilus have been investigated, using two intracellular micro-electrodes, when either Ca ions or Ba ions are the charge carriers. In cells bathed in Ca-free Ba solution the two inward currents, named current I and current II, could be identified and studied in the absence of outward currents. The two inward currents could also be separated by addition of the plant lectin concanavalin A (0.5 microgram/ml) to the external medium, which resulted in the selective inhibition of current I. When the holding potential was set at values between -45 and -65 mV (normal resting potential is -50 mV), current I was shifted parallel to the holding potential along the voltage axis. This shift was 7.6 mV per 10 mV change in holding potential. The amplitude and voltage relationship of current II was not affected by these changes in the holding potential. The amplitude of current I in Ba solution was maximal when the membrane potential was held at -55 mV; it decreased with higher and lower holding potentials. The rate of activation of current I remained virtually unaffected at holding potentials between -45 and -60 mV, and was somewhat reduced at a holding potential of -65 mV. When the extracellular Ca concentration was varied between 0.1 and 5.0 mM, or when the cells were loaded with EGTA to reduce the intracellular level of ionized Ca, the resting membrane potential and the voltage relationships of both current I and II and of the outward current were shifted along the voltage axis according to the expected changes in membrane surface potential. Double-pulse experiments with varying interval potentials suggested voltage-dependent inactivation of current I and Ca-dependent inactivation of current II. Pre-hyperpolarizing steps of only 1 mV amplitude and 30 ms duration could result in the activation of current I, indicating that the activation voltage of current I closely followed the actual membrane potential. Hence, the same voltage steps elicited similar current I amplitudes with holding potentials between -45 and -60 mV. The results indicate that current II displays voltage properties described for Ca channels in other ciliates and many multicellular preparations, while current I shows an unusual voltage behaviour, which might be regarded as an 'adaptive type of excitation'.(ABSTRACT TRUNCATED AT 400 WORDS)