Abstract
We investigated the influence of ion compositions on the membrane potential in LA-N-1 human neuroblastoma cells using bisoxonol as a potential-sensitive fluorescent dye. The ability of K+, ouabain, veratridine, and maitotoxin to induce membrane depolarization was evaluated. Increasing concentrations of K+ ions from 10 to 50 mM caused a dose-dependent increase of bisoxonol fluorescence, which was completely independent on Na+ and Ca2+. Ouabain (5 mM), an inhibitor of the Na+, K+-ATPase, failed to induce membrane depolarization. Veratridine (40 and 100 μM), a Na+ channel activator, only in the presence of 10 μg of Leiurus scorpion venom reduced the membrane potential. Maitotoxin (MTX) from 3 to 10 ng/mL depolarized LA-N-1 cells in a dose-dependent manner, and produced a rapid and sustained increase of intracellular free calcium monitored by means of fluorescent probe fura-2. The MTX-induced depolarization and the increase in cytosolic free calcium concentration were dependent on extracellular Ca2+ ions. On the other hand, Na+ ions also seem to be, although only partially, implicated in the MTX effects, since both the blockade of tetrodotoxin (TTX)-sensitive voltage-operated Na+ channels and the removal of Na+ ions were able to reduce the depolarization. In conclusion, our data indicate that the depolarizing action of MTX on LA-N-1 cells is Ca2+- and Na+-dependent, although the latter only partially, and that this effect is dependent on Ca2+ influx into the cells likely through a voltage-insensitive calcium-entry system.
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Abbreviations
- MTX:
-
maitotoxin
- TTX:
-
tetrodotoxin
- VSCC:
-
voltage-sensitive calcium channel
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Sorrentino, G., Monsurrõ, M.R., Singh, I.N. et al. Membrane depolarization in LA-N-1 cells. Molecular and Chemical Neuropathology 30, 199–211 (1997). https://doi.org/10.1007/BF02815098
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DOI: https://doi.org/10.1007/BF02815098