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Distinct modes of perimembrane TRP channel turnover revealed by TIR-FRAP

Transient Receptor Potential (TRP) channels form a broadly expressed and functionally diverse family of cation channels involved in various (patho)physiological processes. Whereas the mechanisms that control opening of TRP channels have been extensively studied, little is known about the transport p...

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Autores principales: Ghosh, Debapriya, Segal, Andrei, Voets, Thomas
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4236744/
https://www.ncbi.nlm.nih.gov/pubmed/25407951
http://dx.doi.org/10.1038/srep07111
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author Ghosh, Debapriya
Segal, Andrei
Voets, Thomas
author_facet Ghosh, Debapriya
Segal, Andrei
Voets, Thomas
author_sort Ghosh, Debapriya
collection PubMed
description Transient Receptor Potential (TRP) channels form a broadly expressed and functionally diverse family of cation channels involved in various (patho)physiological processes. Whereas the mechanisms that control opening of TRP channels have been extensively studied, little is known about the transport processes of TRP channels to and within the plasma membrane. Here we used Total Internal Reflection - Fluorescence Recovery after Photobleaching (TIR-FRAP) to selectively visualize and bleach the fluorescently labeled TRP channels TRPV2 and TRPM4 in close proximity of the glass-plasma membrane interface, allowing detailed analysis of their perimembrane dynamics. We show that recovery of TRPM4 occurs via 200-nm diameter transport vesicles, and demonstrate the full fusion of such vesicles with the plasma membrane. In contrast, TRPV2 recovery proceeded mainly via lateral diffusion from non-bleached areas of the plasma membrane. Analysis of the two-dimensional channel diffusion kinetics yielded 2D diffusion coefficients ranging between 0.1 and 0.3 μm(2)/s, suggesting that these TRP channels move relatively unrestricted within the plasma membrane. These data demonstrate distinct modes of TRP channel turnover at the plasma membrane and illustrate the usefulness of TIR-FRAP to monitor these processes with high resolution.
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spelling pubmed-42367442014-11-25 Distinct modes of perimembrane TRP channel turnover revealed by TIR-FRAP Ghosh, Debapriya Segal, Andrei Voets, Thomas Sci Rep Article Transient Receptor Potential (TRP) channels form a broadly expressed and functionally diverse family of cation channels involved in various (patho)physiological processes. Whereas the mechanisms that control opening of TRP channels have been extensively studied, little is known about the transport processes of TRP channels to and within the plasma membrane. Here we used Total Internal Reflection - Fluorescence Recovery after Photobleaching (TIR-FRAP) to selectively visualize and bleach the fluorescently labeled TRP channels TRPV2 and TRPM4 in close proximity of the glass-plasma membrane interface, allowing detailed analysis of their perimembrane dynamics. We show that recovery of TRPM4 occurs via 200-nm diameter transport vesicles, and demonstrate the full fusion of such vesicles with the plasma membrane. In contrast, TRPV2 recovery proceeded mainly via lateral diffusion from non-bleached areas of the plasma membrane. Analysis of the two-dimensional channel diffusion kinetics yielded 2D diffusion coefficients ranging between 0.1 and 0.3 μm(2)/s, suggesting that these TRP channels move relatively unrestricted within the plasma membrane. These data demonstrate distinct modes of TRP channel turnover at the plasma membrane and illustrate the usefulness of TIR-FRAP to monitor these processes with high resolution. Nature Publishing Group 2014-11-19 /pmc/articles/PMC4236744/ /pubmed/25407951 http://dx.doi.org/10.1038/srep07111 Text en Copyright © 2014, Macmillan Publishers Limited. All rights reserved http://creativecommons.org/licenses/by-nc-sa/4.0/ This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder in order to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/4.0/
spellingShingle Article
Ghosh, Debapriya
Segal, Andrei
Voets, Thomas
Distinct modes of perimembrane TRP channel turnover revealed by TIR-FRAP
title Distinct modes of perimembrane TRP channel turnover revealed by TIR-FRAP
title_full Distinct modes of perimembrane TRP channel turnover revealed by TIR-FRAP
title_fullStr Distinct modes of perimembrane TRP channel turnover revealed by TIR-FRAP
title_full_unstemmed Distinct modes of perimembrane TRP channel turnover revealed by TIR-FRAP
title_short Distinct modes of perimembrane TRP channel turnover revealed by TIR-FRAP
title_sort distinct modes of perimembrane trp channel turnover revealed by tir-frap
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4236744/
https://www.ncbi.nlm.nih.gov/pubmed/25407951
http://dx.doi.org/10.1038/srep07111
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