The Journal of General Physiology
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Published online Feb 27 2006. doi:10.1085/jgp.200509402
The Rockefeller University Press, 0022-1295 $8.00
JGP, Volume 127, Number 3, 329-340
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ARTICLE

 On the Interaction of Neomycin with the Slow Vacuolar Channel of Arabidopsis thaliana



Joachim Scholz-Starke, Armando Carpaneto, and Franco Gambale

Istituto di Biofisica, Consiglio Nazionale delle Ricerche, 16149 Genoa, Italy

Correspondence to Joachim Scholz-Starke: scholz{at}ge.ibf.cnr.it

This study investigates the interaction of the aminoglycoside antibiotic neomycin with the slow vacuolar (SV) channel in vacuoles from Arabidopsis thaliana mesophyll cells. Patch-clamp experiments in the excised patch configuration revealed a complex pattern of neomycin effects on the channel: applied at concentrations in the submicromolar to millimolar range neomycin (a) blocked macroscopic SV currents in a voltage- and concentration-dependent manner, (b) slowed down activation and deactivation kinetics of the channel, and most interestingly, (c) at concentrations above 10 µM, neomycin shifted the SV activation threshold towards negative membrane potentials, causing a two-phasic activation at high concentrations. Single channel experiments showed that neomycin causes these macroscopic effects by combining a decrease of the single channel conductance with a concomitant increase of the channel's open probability. Our results clearly demonstrate that the SV channel can be activated at physiologically relevant tonoplast potentials in the presence of an organic effector molecule. We therefore propose the existence of a cellular equivalent regulating the activity of the SV channel in vivo.

Abbreviations used in this paper: DTT, dithiothreitol; IP3, inositol 1,4,5-triphosphate; RyR, ryanodine receptor; SV, slow vacuolar.


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