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Short-Term Regulation of Murine Colonic NBCe1-B (Electrogenic Na(+)/HCO(3) (−) Cotransporter) Membrane Expression and Activity by Protein Kinase C
The colonic mucosa actively secretes HCO(3) (−), and several lines of evidence point to an important role of Na(+)/HCO(3) (−) cotransport (NBC) as a basolateral HCO(3) (−) import pathway. We could recently demonstrate that the predominant NBC isoform in murine colonic crypts is electrogenic NBCe1-B,...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3958514/ https://www.ncbi.nlm.nih.gov/pubmed/24642792 http://dx.doi.org/10.1371/journal.pone.0092275 |
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author | May, Oliver Yu, Haoyang Riederer, Brigitte Manns, Michael P. Seidler, Ursula Bachmann, Oliver |
author_facet | May, Oliver Yu, Haoyang Riederer, Brigitte Manns, Michael P. Seidler, Ursula Bachmann, Oliver |
author_sort | May, Oliver |
collection | PubMed |
description | The colonic mucosa actively secretes HCO(3) (−), and several lines of evidence point to an important role of Na(+)/HCO(3) (−) cotransport (NBC) as a basolateral HCO(3) (−) import pathway. We could recently demonstrate that the predominant NBC isoform in murine colonic crypts is electrogenic NBCe1-B, and that secretagogues cause NBCe1 exocytosis, which likely represents a component of NBC activation. Since protein kinase C (PKC) plays a key role in the regulation of ion transport by trafficking events, we asked whether it is also involved in the observed NBC activity increase. Crypts were isolated from murine proximal colon to assess PKC activation as well as NBC function and membrane abundance using fluorometric pH(i) measurements and cell surface biotinylation, respectively. PKC isoform translocation and phosphorylation occurred in response to PMA-, as well as secretagogue stimulation. The conventional and novel PKC inhibitors Gö6976 or Gö6850 did not alter NBC function or surface expression by themselves, but stimulation with forskolin (10(−5) M) or carbachol (10(−4) M) in their presence led to a significant decrease in NBC-mediated proton flux, and biotinylated NBCe1. Our data thus indicate that secretagogues lead to PKC translocation and phosphorylation in murine colonic crypts, and that PKC is necessary for the increase in NBC transport rate and membrane abundance caused by cholinergic and cAMP-dependent stimuli. |
format | Online Article Text |
id | pubmed-3958514 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-39585142014-03-24 Short-Term Regulation of Murine Colonic NBCe1-B (Electrogenic Na(+)/HCO(3) (−) Cotransporter) Membrane Expression and Activity by Protein Kinase C May, Oliver Yu, Haoyang Riederer, Brigitte Manns, Michael P. Seidler, Ursula Bachmann, Oliver PLoS One Research Article The colonic mucosa actively secretes HCO(3) (−), and several lines of evidence point to an important role of Na(+)/HCO(3) (−) cotransport (NBC) as a basolateral HCO(3) (−) import pathway. We could recently demonstrate that the predominant NBC isoform in murine colonic crypts is electrogenic NBCe1-B, and that secretagogues cause NBCe1 exocytosis, which likely represents a component of NBC activation. Since protein kinase C (PKC) plays a key role in the regulation of ion transport by trafficking events, we asked whether it is also involved in the observed NBC activity increase. Crypts were isolated from murine proximal colon to assess PKC activation as well as NBC function and membrane abundance using fluorometric pH(i) measurements and cell surface biotinylation, respectively. PKC isoform translocation and phosphorylation occurred in response to PMA-, as well as secretagogue stimulation. The conventional and novel PKC inhibitors Gö6976 or Gö6850 did not alter NBC function or surface expression by themselves, but stimulation with forskolin (10(−5) M) or carbachol (10(−4) M) in their presence led to a significant decrease in NBC-mediated proton flux, and biotinylated NBCe1. Our data thus indicate that secretagogues lead to PKC translocation and phosphorylation in murine colonic crypts, and that PKC is necessary for the increase in NBC transport rate and membrane abundance caused by cholinergic and cAMP-dependent stimuli. Public Library of Science 2014-03-18 /pmc/articles/PMC3958514/ /pubmed/24642792 http://dx.doi.org/10.1371/journal.pone.0092275 Text en © 2014 May 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 May, Oliver Yu, Haoyang Riederer, Brigitte Manns, Michael P. Seidler, Ursula Bachmann, Oliver Short-Term Regulation of Murine Colonic NBCe1-B (Electrogenic Na(+)/HCO(3) (−) Cotransporter) Membrane Expression and Activity by Protein Kinase C |
title | Short-Term Regulation of Murine Colonic NBCe1-B (Electrogenic Na(+)/HCO(3)
(−) Cotransporter) Membrane Expression and Activity by Protein Kinase C |
title_full | Short-Term Regulation of Murine Colonic NBCe1-B (Electrogenic Na(+)/HCO(3)
(−) Cotransporter) Membrane Expression and Activity by Protein Kinase C |
title_fullStr | Short-Term Regulation of Murine Colonic NBCe1-B (Electrogenic Na(+)/HCO(3)
(−) Cotransporter) Membrane Expression and Activity by Protein Kinase C |
title_full_unstemmed | Short-Term Regulation of Murine Colonic NBCe1-B (Electrogenic Na(+)/HCO(3)
(−) Cotransporter) Membrane Expression and Activity by Protein Kinase C |
title_short | Short-Term Regulation of Murine Colonic NBCe1-B (Electrogenic Na(+)/HCO(3)
(−) Cotransporter) Membrane Expression and Activity by Protein Kinase C |
title_sort | short-term regulation of murine colonic nbce1-b (electrogenic na(+)/hco(3)
(−) cotransporter) membrane expression and activity by protein kinase c |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3958514/ https://www.ncbi.nlm.nih.gov/pubmed/24642792 http://dx.doi.org/10.1371/journal.pone.0092275 |
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