Sexual behaviour in Euplotes raikovi is accompanied by pheromone-induced modifications of ionic currents.

In the marine ciliate Euplotes raikovi, pheromone released by a complementary mating type (nonself pheromone) induces typical sexual behaviour, whereas self pheromone released by the same mating type generally has no effect. Nonself pheromone evokes a reduction of the mean walking speed by 66 %, a threefold increase in the frequency and duration of long-lasting rest phases and a doubling in the number of side-stepping reactions. Consequently, translocation is strongly reduced and the cells remain in a small area. This could increase the probability of finding a sexual partner for pair formation (conjugation). The usual pattern of rhythmic, spontaneous depolarizations controlling the walking rhythm is absent in nonself-pheromone-stimulated cells. The remaining depolarizations arise from a 4 mV hyperpolarized membrane potential and do not reach the usual amplitudes of 15-20 mV but only of 6-10 mV. In addition, the amplitudes of K+ currents are increased at depolarizations of more than 20 mV by at least 30 %. Hyperpolarization- and depolarization-activated Na+ current amplitudes are increased, whereas the Ca2+ current amplitude remains nearly unaffected.

Calcium-dependent transient potassium outward current in the marine ciliate Euplotes vannus.

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.

Calcium-dependent sodium current in the marine ciliate Euplotes vannus.

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+.

Membrane excitability and membrane currents in the marine ciliate Euplotes vannus.

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+).

Transport of choline by membrane vesicles prepared from synaptosomes of insect nervous tissue.

Membrane vesicles derived from synaptic plasma membranes have been isolated from insect nervous tissue. High affinity uptake of choline into these vesicles has been demonstrated using artificially imposed electrochemical gradients as the sole energy source. The transport of choline is strictly dependent on the presence of Na(+) and Cl(?) in the external medium and is mainly driven by a Na(+) gradient. Inhibition by proton ionophores and stimulation by valinomycin suggest that choline uptake is an electrogenic process which is optimal in the presence of a membrane potential. In addition, the process is inhibited by alkaloid neurotoxins veratridine and aconitine; this inhibitory effect is prevented by tetrodotoxin. The data are consistent with the predominant role of ion chemical gradients and an electrical membrane potential in energizing the uptake of choline.