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Characterization of the Ca(2+)-Gated and Voltage-Dependent K(+)-Channel Slo-1 of Nematodes and Its Interaction with Emodepside

The cyclooctadepsipeptide emodepside and its parent compound PF1022A are broad-spectrum nematicidal drugs which are able to eliminate nematodes resistant to other anthelmintics. The mode of action of cyclooctadepsipeptides is only partially understood, but involves the latrophilin Lat-1 receptor and...

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Autores principales: Kulke, Daniel, von Samson-Himmelstjerna, Georg, Miltsch, Sandra M., Wolstenholme, Adrian J., Jex, Aaron R., Gasser, Robin B., Ballesteros, Cristina, Geary, Timothy G., Keiser, Jennifer, Townson, Simon, Harder, Achim, Krücken, Jürgen
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4270693/
https://www.ncbi.nlm.nih.gov/pubmed/25521608
http://dx.doi.org/10.1371/journal.pntd.0003401
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author Kulke, Daniel
von Samson-Himmelstjerna, Georg
Miltsch, Sandra M.
Wolstenholme, Adrian J.
Jex, Aaron R.
Gasser, Robin B.
Ballesteros, Cristina
Geary, Timothy G.
Keiser, Jennifer
Townson, Simon
Harder, Achim
Krücken, Jürgen
author_facet Kulke, Daniel
von Samson-Himmelstjerna, Georg
Miltsch, Sandra M.
Wolstenholme, Adrian J.
Jex, Aaron R.
Gasser, Robin B.
Ballesteros, Cristina
Geary, Timothy G.
Keiser, Jennifer
Townson, Simon
Harder, Achim
Krücken, Jürgen
author_sort Kulke, Daniel
collection PubMed
description The cyclooctadepsipeptide emodepside and its parent compound PF1022A are broad-spectrum nematicidal drugs which are able to eliminate nematodes resistant to other anthelmintics. The mode of action of cyclooctadepsipeptides is only partially understood, but involves the latrophilin Lat-1 receptor and the voltage- and calcium-activated potassium channel Slo-1. Genetic evidence suggests that emodepside exerts its anthelmintic activity predominantly through Slo-1. Indeed, slo-1 deficient Caenorhabditis elegans strains are completely emodepside resistant. However, direct effects of emodepside on Slo-1 have not been reported and these channels have only been characterized for C. elegans and related Strongylida. Molecular and bioinformatic analyses identified full-length Slo-1 cDNAs of Ascaris suum, Parascaris equorum, Toxocara canis, Dirofilaria immitis, Brugia malayi, Onchocerca gutturosa and Strongyloides ratti. Two paralogs were identified in the trichocephalids Trichuris muris, Trichuris suis and Trichinella spiralis. Several splice variants encoding truncated channels were identified in Trichuris spp. Slo-1 channels of trichocephalids form a monophyletic group, showing that duplication occurred after the divergence of Enoplea and Chromadorea. To explore the function of a representative protein, C. elegans Slo-1a was expressed in Xenopus laevis oocytes and studied in electrophysiological (voltage-clamp) experiments. Incubation of oocytes with 1-10 µM emodepside caused significantly increased currents over a wide range of step potentials in the absence of experimentally increased intracellular Ca(2+), suggesting that emodepside directly opens C. elegans Slo-1a. Emodepside wash-out did not reverse the effect and the Slo-1 inhibitor verruculogen was only effective when applied before, but not after, emodepside. The identification of several splice variants and paralogs in some parasitic nematodes suggests that there are substantial differences in channel properties among species. Most importantly, this study showed for the first time that emodepside directly opens a Slo-1 channel, significantly improving the understanding of the mode of action of this drug class.
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spelling pubmed-42706932014-12-26 Characterization of the Ca(2+)-Gated and Voltage-Dependent K(+)-Channel Slo-1 of Nematodes and Its Interaction with Emodepside Kulke, Daniel von Samson-Himmelstjerna, Georg Miltsch, Sandra M. Wolstenholme, Adrian J. Jex, Aaron R. Gasser, Robin B. Ballesteros, Cristina Geary, Timothy G. Keiser, Jennifer Townson, Simon Harder, Achim Krücken, Jürgen PLoS Negl Trop Dis Research Article The cyclooctadepsipeptide emodepside and its parent compound PF1022A are broad-spectrum nematicidal drugs which are able to eliminate nematodes resistant to other anthelmintics. The mode of action of cyclooctadepsipeptides is only partially understood, but involves the latrophilin Lat-1 receptor and the voltage- and calcium-activated potassium channel Slo-1. Genetic evidence suggests that emodepside exerts its anthelmintic activity predominantly through Slo-1. Indeed, slo-1 deficient Caenorhabditis elegans strains are completely emodepside resistant. However, direct effects of emodepside on Slo-1 have not been reported and these channels have only been characterized for C. elegans and related Strongylida. Molecular and bioinformatic analyses identified full-length Slo-1 cDNAs of Ascaris suum, Parascaris equorum, Toxocara canis, Dirofilaria immitis, Brugia malayi, Onchocerca gutturosa and Strongyloides ratti. Two paralogs were identified in the trichocephalids Trichuris muris, Trichuris suis and Trichinella spiralis. Several splice variants encoding truncated channels were identified in Trichuris spp. Slo-1 channels of trichocephalids form a monophyletic group, showing that duplication occurred after the divergence of Enoplea and Chromadorea. To explore the function of a representative protein, C. elegans Slo-1a was expressed in Xenopus laevis oocytes and studied in electrophysiological (voltage-clamp) experiments. Incubation of oocytes with 1-10 µM emodepside caused significantly increased currents over a wide range of step potentials in the absence of experimentally increased intracellular Ca(2+), suggesting that emodepside directly opens C. elegans Slo-1a. Emodepside wash-out did not reverse the effect and the Slo-1 inhibitor verruculogen was only effective when applied before, but not after, emodepside. The identification of several splice variants and paralogs in some parasitic nematodes suggests that there are substantial differences in channel properties among species. Most importantly, this study showed for the first time that emodepside directly opens a Slo-1 channel, significantly improving the understanding of the mode of action of this drug class. Public Library of Science 2014-12-18 /pmc/articles/PMC4270693/ /pubmed/25521608 http://dx.doi.org/10.1371/journal.pntd.0003401 Text en © 2014 Kulke et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Kulke, Daniel
von Samson-Himmelstjerna, Georg
Miltsch, Sandra M.
Wolstenholme, Adrian J.
Jex, Aaron R.
Gasser, Robin B.
Ballesteros, Cristina
Geary, Timothy G.
Keiser, Jennifer
Townson, Simon
Harder, Achim
Krücken, Jürgen
Characterization of the Ca(2+)-Gated and Voltage-Dependent K(+)-Channel Slo-1 of Nematodes and Its Interaction with Emodepside
title Characterization of the Ca(2+)-Gated and Voltage-Dependent K(+)-Channel Slo-1 of Nematodes and Its Interaction with Emodepside
title_full Characterization of the Ca(2+)-Gated and Voltage-Dependent K(+)-Channel Slo-1 of Nematodes and Its Interaction with Emodepside
title_fullStr Characterization of the Ca(2+)-Gated and Voltage-Dependent K(+)-Channel Slo-1 of Nematodes and Its Interaction with Emodepside
title_full_unstemmed Characterization of the Ca(2+)-Gated and Voltage-Dependent K(+)-Channel Slo-1 of Nematodes and Its Interaction with Emodepside
title_short Characterization of the Ca(2+)-Gated and Voltage-Dependent K(+)-Channel Slo-1 of Nematodes and Its Interaction with Emodepside
title_sort characterization of the ca(2+)-gated and voltage-dependent k(+)-channel slo-1 of nematodes and its interaction with emodepside
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4270693/
https://www.ncbi.nlm.nih.gov/pubmed/25521608
http://dx.doi.org/10.1371/journal.pntd.0003401
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