No abstract

Among secondary metabolites, the acetylated hemiacetal sesquiterpene euplotin C has been isolated from the marine, ciliated protist Euplotes crassus, and provides an effective mechanism for reducing populations of potential competitors through its cytotoxic properties. However, intracellular signaling mechanisms and their functional correlates mediating the ecological role of euplotin C are largely unknown. We report here that, in E. vannus (an Euplotes morphospecies that does not produce euplotin C and shares with E. crasssus the same interstitial habitat), euplotin C rapidly increases the intracellular concentration of both Ca(2+) and Na(+), suggesting a generalized effect of this metabolite on cation transport systems. In addition, euplotin C does not induce oxidative stress, but modulates the electrical properties of E. vannus through an increase of the amplitude of graded action potentials. These events parallel the disassembling of the ciliary structures, the inhibition of cell motility, the occurrence of aberrant cytoplasmic vacuoles, and the rapid inhibition of phagocytic activity. Euplotin C also increases lysosomal pH and decreases lysosomal membrane stability of E. vannus. These results suggest that euplotin C exerts a marked disruption of those homeostatic mechanisms whose efficiency represents the essential prerequisite to face the challenges of the interstitial environment.

No abstract

Ionic currents were recorded in cultured oligodendrocytes from the brain of trout using the whole-cell configuration of the patch-clamp technique. Outward currents were evoked at membrane potentials more positive than -40 mV, which could be separated into two components according to their kinetic parameters and their sensitivity to the holding potential: a fast inactivating current which was completely suppressed by 4-aminopyridine and reduced by tetraethylammonium and a slow steady-state conductance which was similarly sensitive to both potassium channel blockers. The current reversal potential was close to the potassium equilibrium potential. In contrast to mammalian oligodendrocytes but in similarity with cultured Schwann cells, trout oligodendrocytes did not exhibit any inwardly rectifying currents at hyperpolarized membrane potentials.

Voltage-gated ionic currents were recorded from cultured trout astrocytes with the whole-cell variation of the patch-clamp technique. In a subpopulation of astrocytes depolarizations above -40 mV activated a fast transient inward current that was identified as a sodium current by ion substitution experiments, its current reversal potential, and its TTX-sensitivity. Regarding threshold of activation, peak current voltage, and amplitude this current closely resembled those previously described for mammalian astrocytes. Voltage-dependence of inactivation and kinetics, however, markedly differed from the "glial-like" sodium current occurring in mammalian hippocampal or optic nerve astrocytes, since the sodium current of trout astrocytes exhibited a faster time course of activation and decay and a more depolarized steady-state inactivation curve with midpoints close to -60 mV. During a period of 2 weeks in culture the biophysical properties of the sodium current did not change significantly, albeit a continuous decrease in current density was observed. At depolarizing voltage steps positive to -40 mV, additionally voltage-gated potassium outward currents were evoked, which could be separated into a steady-state current with delayed rectifier properties and an inactivating component resembling the A-type current. Moreover, in a subpopulation of astrocytes an inward potassium current was elicited at hyperpolarizing potentials, which exhibited biophysical features consistent with the potassium inward rectifier of mammalian astrocytes.