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Electrical responses upon mechanostimulation at the posterior cell end were investigated in the marine hypotrichous ciliate Euplotes vannus. A new mechanostimulator was developed to mimic stimuli that are identical with those involved in cell-cell collisions. The receptor potential hyperpolarized by 18-35 mV within 12-25 msec, reached a peak value of -62 mV with a delay of 4-9 msec after membrane deformation, and was deactivated after 50-70 msec. Cirri were stimulated to beat accelerated backward. The corresponding receptor current exerted a similar time course with a peak of 2.4 nA. The shift of the reversal potential by 57.6 mV at a tenfold increase of [K+]o identifies potassium ions as current carriers within the development of the receptor potential. An intracellular K concentration of 355 mmol/liter was calculated for cells in a medium that was composed similar to sea-water. The mechanically activated potassium current was totally inhibited by extracellular TEA and intracellular Cs+, and partially inhibited by extracellular 4-AP. The total inhibition of the current by injected EGTA points to a Ca dependence of the posterior mechanosensitivity. It was confirmed by the increase of the peak current amplitude with rising [Ca2+]o. Sodium presumably repolarizes the receptor potential because the repolarization was delayed and after-depolarizations were eliminated in media without sodium. Since deciliation did not affect mechanosensitivity, the corresponding ion channels reside within the soma membrane. [ABSTRACT FROM AUTHOR]

The ionic mechanisms underlying inward or anomalous rectification have been studied in the marine hypotrichous ciliate Euplotes vannus. Inward-current pulses of moderate amplitude elicited time-dependent rectification that started from a hyperpolarization peak and was expressed as a depolarizing sag towards rest. Voltage-clamp analysis showed that this depolarization is caused by the activation of a complex inward current that does not inactivate with time. The current is carried by a major Na and a minor K component. The Na-current component has been identified by its concentration-dependent reduction in low extracellular Na solutions and the capability of Li+ as Na substitute to carry the current, though with a slightly reduced amplitude. The K-current component has been isolated from the total current after the replacement of Na+ within the experimental solution. It was blocked in media that contained 10 mmol/liter TEA, a well-known blocker for K inwardly rectifying currents. TEA was only effective at membrane potentials close to or negative to the potassium equilibrium potential. The inward current was reduced after the injection of the Ca chelator EGTA into the cell. Also the elimination of the ciliary membrane, by deciliating cells with ethanol, reduced the amplitude of the inwardly rectifying currents. Both experiments indicate a regulatory function of Ca2+ in inward rectification. [ABSTRACT FROM AUTHOR]

Ca and Na inward currents were recorded upon depolarizations in Euplotes after the blockage of K outward currents with intracellular Cs ions. The Na current was analyzed under voltage clamp and had the following properties: it activated to a maximum within 150 msec and partly inactivated during sustained voltage steps. It had a positive equilibrium potential between 25 and 30 mV and could be carried by Na or Li ions but not by K, choline or Tris ions. The current revealed a prominent associated inward tail current which deactivated with a single-exponential time constant of 118 msec. Both the current and its tail were strongly reduced after reduction of the extracellular Na concentration. Externally applied K channel blocker tetraethylammonium chloride did not block the current. Either EGTA injection into the cell or nonlethal deciliation with ethanol eliminated the current and its tail. These results indicate the existence of a Na conductance within the membrane of Euplotes which is activated by the intracellular level of free Ca2+. [ABSTRACT FROM AUTHOR]

We have isolated the early Ca current (ICa) from the whole cell current that activates upon depolarisations in the marine ciliate Euplotes vannus. The peak of ICa activated within 4.2 ms at depolarisations to 5 mV with an amplitude of 2.5 +/- 0.35 nA and was reduced to 1.0 +/- 0.14 nA (n = 5) when the extracellular Ca concentration was changed from 10 to 1 mM. The voltage-dependent activation curve was steeper and shifted to more negative values when external Ca2+ was replaced by Ba2+. The early inward current inactivated with a double-exponential time course including a fast and a slow component, and no inactivation was recorded with Ba2+. The time constants for the recovery from inactivation varied between 44 and 153 ms according to the depolarisation-dependent Ca influx. At the common resting potential of -25 mV, ICa was not steady-state inactivated; ICa half-inactivated at -14.5 mV, and totally inactivated at -5 mV. ICa was inhibited by 10 mM extracellular Cd2+. The peptides omega-conotoxin-GVIA (20 microM), omega-conotoxin-MVIIC (600 nM), omega-agatoxin-IVA (60 nM) and calciseptine (900 nM) did not block ICa. The benzothiazepine-derivative diltiazem (100 microM) and the dihydropiridine nifedipine (100 microM) inhibited 51% and 33% of ICa, respectively. The naphthalene sulfonamide W7 reduced ICa with an inhibition coefficient of 33 microM.

In the marine hypotrichous ciliate Euplotes vannus, the transient K+ outward current, IK fast, was studied by use of a single-microelectrode voltage-clamp equipment. Activation and inactivation kinetics, and steady-state inactivation are comparable to the properties of A-currents. Not typical for this type of current is its insensitivity to either 4-AP or 3,4-AP and its Ca2+ dependence which was derived from its inhibition by either extracellular Cd2+, La3+, D-600, or by intracellular BAPTA. Actual amplitudes of IK fast were obtained from a composite current, by subtraction of early parts of a slowly activating K+ current, IK slow, and of the early, transient Ca2+ inward current, ICa fast, that is typical for ciliates. IK fast counteracts ICa fast during the first milliseconds after onset of depolarization such that the composite current is purely outward directed.

Electrical properties of E. vannus were investigated by use of constant current injection, voltage-clamp, and isoosmotic ion substitution. The resting potential of approximately -40 mV was K(+) and Ca(2+)-dependent. Spontaneous depolarizations occurred frequently with peaks around -20 mV and durations from several hundred ms to several s. External Ba(2+) or internal Cs(+) induced all-or-none action potentials. Current stimuli induced Ca(2+)-dependent graded action potentials. Sr(2+) or Ba(2+), but not Mg(2+), instead of Ca(2+) increased the regenerative response. Repolarization occurred in two steps: a first fast and a second slow one. It was strongly modified by the Ca(2+) substitutes. A voltage-dependent small Ca(2+) inward current was activated at depolarizations beyond -20 mV. It triggered a fast and a slowly activating K(+) outward current and was itself short-circuited by the fast K(+) current. Therefore, it could only be measured when K(+) currents were not activated or inhibited. A slowly activating Na(+) inward current was identified that turned to outward direction after replacement of external Na(+) by choline(+). The K(+) outward currents differed in their sensitivity to external TEA(+) and in their inactivation kinetics. All currents were correlated to the voltage-dependent influx of Ca(2+)., (Copyright © 1988 Gustav Fischer Verlag · Stuttgart · New York. Published by Elsevier GmbH.. All rights reserved.)