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Dynamics of Kv1 Channel Transport in Axons
Concerted actions of various ion channels that are precisely targeted along axons are crucial for action potential initiation and propagation, and neurotransmitter release. However, the dynamics of channel protein transport in axons remain unknown. Here, using time-lapse imaging, we found fluorescen...
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Formato: | Texto |
Lenguaje: | English |
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Public Library of Science
2010
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2915926/ https://www.ncbi.nlm.nih.gov/pubmed/20694152 http://dx.doi.org/10.1371/journal.pone.0011931 |
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author | Gu, Yuanzheng Gu, Chen |
author_facet | Gu, Yuanzheng Gu, Chen |
author_sort | Gu, Yuanzheng |
collection | PubMed |
description | Concerted actions of various ion channels that are precisely targeted along axons are crucial for action potential initiation and propagation, and neurotransmitter release. However, the dynamics of channel protein transport in axons remain unknown. Here, using time-lapse imaging, we found fluorescently tagged Kv1.2 voltage-gated K(+) channels (YFP-Kv1.2) moved bi-directionally in discrete puncta along hippocampal axons. Expressing Kvβ2, a Kv1 accessory subunit, markedly increased the velocity, the travel distance, and the percentage of moving time of these puncta in both anterograde and retrograde directions. Suppressing the Kvβ2-associated protein, plus-end binding protein EB1 or kinesin II/KIF3A, by siRNA, significantly decreased the velocity of YFP-Kv1.2 moving puncta in both directions. Kvβ2 mutants with disrupted either Kv1.2-Kvβ2 binding or Kvβ2-EB1 binding failed to increase the velocity of YFP-Kv1.2 puncta, confirming a central role of Kvβ2. Furthermore, fluorescently tagged Kv1.2 and Kvβ2 co-moved along axons. Surprisingly, when co-moving with Kv1.2 and Kvβ2, EB1 appeared to travel markedly faster than its plus-end tracking. Finally, using fission yeast S. pombe expressing YFP-fusion proteins as reference standards to calibrate our microscope, we estimated the numbers of YFP-Kv1.2 tetramers in axonal puncta. Taken together, our results suggest that proper amounts of Kv1 channels and their associated proteins are required for efficient transport of Kv1 channel proteins along axons. |
format | Text |
id | pubmed-2915926 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2010 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-29159262010-08-05 Dynamics of Kv1 Channel Transport in Axons Gu, Yuanzheng Gu, Chen PLoS One Research Article Concerted actions of various ion channels that are precisely targeted along axons are crucial for action potential initiation and propagation, and neurotransmitter release. However, the dynamics of channel protein transport in axons remain unknown. Here, using time-lapse imaging, we found fluorescently tagged Kv1.2 voltage-gated K(+) channels (YFP-Kv1.2) moved bi-directionally in discrete puncta along hippocampal axons. Expressing Kvβ2, a Kv1 accessory subunit, markedly increased the velocity, the travel distance, and the percentage of moving time of these puncta in both anterograde and retrograde directions. Suppressing the Kvβ2-associated protein, plus-end binding protein EB1 or kinesin II/KIF3A, by siRNA, significantly decreased the velocity of YFP-Kv1.2 moving puncta in both directions. Kvβ2 mutants with disrupted either Kv1.2-Kvβ2 binding or Kvβ2-EB1 binding failed to increase the velocity of YFP-Kv1.2 puncta, confirming a central role of Kvβ2. Furthermore, fluorescently tagged Kv1.2 and Kvβ2 co-moved along axons. Surprisingly, when co-moving with Kv1.2 and Kvβ2, EB1 appeared to travel markedly faster than its plus-end tracking. Finally, using fission yeast S. pombe expressing YFP-fusion proteins as reference standards to calibrate our microscope, we estimated the numbers of YFP-Kv1.2 tetramers in axonal puncta. Taken together, our results suggest that proper amounts of Kv1 channels and their associated proteins are required for efficient transport of Kv1 channel proteins along axons. Public Library of Science 2010-08-04 /pmc/articles/PMC2915926/ /pubmed/20694152 http://dx.doi.org/10.1371/journal.pone.0011931 Text en Gu, Gu. 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 Gu, Yuanzheng Gu, Chen Dynamics of Kv1 Channel Transport in Axons |
title | Dynamics of Kv1 Channel Transport in Axons |
title_full | Dynamics of Kv1 Channel Transport in Axons |
title_fullStr | Dynamics of Kv1 Channel Transport in Axons |
title_full_unstemmed | Dynamics of Kv1 Channel Transport in Axons |
title_short | Dynamics of Kv1 Channel Transport in Axons |
title_sort | dynamics of kv1 channel transport in axons |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2915926/ https://www.ncbi.nlm.nih.gov/pubmed/20694152 http://dx.doi.org/10.1371/journal.pone.0011931 |
work_keys_str_mv | AT guyuanzheng dynamicsofkv1channeltransportinaxons AT guchen dynamicsofkv1channeltransportinaxons |