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Leucine‐rich repeat containing protein LRRC8A is essential for swelling‐activated Cl(−) currents and embryonic development in zebrafish
A volume‐regulated anion channel (VRAC) has been electrophysiologically characterized in innumerable mammalian cell types. VRAC is activated by cell swelling and mediates the volume regulatory efflux of Cl(−) and small organic solutes from cells. Two groups recently identified the mammalian leucine‐...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5064130/ https://www.ncbi.nlm.nih.gov/pubmed/27688432 http://dx.doi.org/10.14814/phy2.12940 |
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author | Yamada, Toshiki Wondergem, Robert Morrison, Rebecca Yin, Viravuth P. Strange, Kevin |
author_facet | Yamada, Toshiki Wondergem, Robert Morrison, Rebecca Yin, Viravuth P. Strange, Kevin |
author_sort | Yamada, Toshiki |
collection | PubMed |
description | A volume‐regulated anion channel (VRAC) has been electrophysiologically characterized in innumerable mammalian cell types. VRAC is activated by cell swelling and mediates the volume regulatory efflux of Cl(−) and small organic solutes from cells. Two groups recently identified the mammalian leucine‐rich repeat containing protein LRRC8A as an essential VRAC component. LRRC8A must be coexpressed with at least one of the other four members of this gene family, LRRC8B‐E, to reconstitute VRAC activity in LRRC8 (−/−) cells. LRRC8 genes likely arose with the origin of chordates. We identified LRRC8A and LRRC8C‐E orthologs in the zebrafish genome and demonstrate that zebrafish embryo cells and differentiated adult cell types express a swelling‐activated Cl(−) current indistinguishable from mammalian VRAC currents. Embryo cell VRAC currents are virtually eliminated by morpholino knockdown of the zebrafish LRRC8A ortholog lrrc8aa. VRAC activity is fully reconstituted in LRRC8 (−/−) human cells by coexpression of zebrafish lrrc8aa and human LRRC8C cDNAs. lrrc8aa expression varies during zebrafish embryogenesis and lrrc8aa knockdown causes pericardial edema and defects in trunk elongation and somatogenesis. Our studies provide confirmation of the importance of LRRC8A in VRAC activity and establish the zebrafish as a model system for characterizing the molecular regulation and physiological roles of VRAC and LRRC8 proteins. |
format | Online Article Text |
id | pubmed-5064130 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-50641302016-10-24 Leucine‐rich repeat containing protein LRRC8A is essential for swelling‐activated Cl(−) currents and embryonic development in zebrafish Yamada, Toshiki Wondergem, Robert Morrison, Rebecca Yin, Viravuth P. Strange, Kevin Physiol Rep Original Research A volume‐regulated anion channel (VRAC) has been electrophysiologically characterized in innumerable mammalian cell types. VRAC is activated by cell swelling and mediates the volume regulatory efflux of Cl(−) and small organic solutes from cells. Two groups recently identified the mammalian leucine‐rich repeat containing protein LRRC8A as an essential VRAC component. LRRC8A must be coexpressed with at least one of the other four members of this gene family, LRRC8B‐E, to reconstitute VRAC activity in LRRC8 (−/−) cells. LRRC8 genes likely arose with the origin of chordates. We identified LRRC8A and LRRC8C‐E orthologs in the zebrafish genome and demonstrate that zebrafish embryo cells and differentiated adult cell types express a swelling‐activated Cl(−) current indistinguishable from mammalian VRAC currents. Embryo cell VRAC currents are virtually eliminated by morpholino knockdown of the zebrafish LRRC8A ortholog lrrc8aa. VRAC activity is fully reconstituted in LRRC8 (−/−) human cells by coexpression of zebrafish lrrc8aa and human LRRC8C cDNAs. lrrc8aa expression varies during zebrafish embryogenesis and lrrc8aa knockdown causes pericardial edema and defects in trunk elongation and somatogenesis. Our studies provide confirmation of the importance of LRRC8A in VRAC activity and establish the zebrafish as a model system for characterizing the molecular regulation and physiological roles of VRAC and LRRC8 proteins. John Wiley and Sons Inc. 2016-09-28 /pmc/articles/PMC5064130/ /pubmed/27688432 http://dx.doi.org/10.14814/phy2.12940 Text en © 2016 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of the American Physiological Society and The Physiological Society. This is an open access article under the terms of the Creative Commons Attribution (http://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Original Research Yamada, Toshiki Wondergem, Robert Morrison, Rebecca Yin, Viravuth P. Strange, Kevin Leucine‐rich repeat containing protein LRRC8A is essential for swelling‐activated Cl(−) currents and embryonic development in zebrafish |
title | Leucine‐rich repeat containing protein LRRC8A is essential for swelling‐activated Cl(−) currents and embryonic development in zebrafish |
title_full | Leucine‐rich repeat containing protein LRRC8A is essential for swelling‐activated Cl(−) currents and embryonic development in zebrafish |
title_fullStr | Leucine‐rich repeat containing protein LRRC8A is essential for swelling‐activated Cl(−) currents and embryonic development in zebrafish |
title_full_unstemmed | Leucine‐rich repeat containing protein LRRC8A is essential for swelling‐activated Cl(−) currents and embryonic development in zebrafish |
title_short | Leucine‐rich repeat containing protein LRRC8A is essential for swelling‐activated Cl(−) currents and embryonic development in zebrafish |
title_sort | leucine‐rich repeat containing protein lrrc8a is essential for swelling‐activated cl(−) currents and embryonic development in zebrafish |
topic | Original Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5064130/ https://www.ncbi.nlm.nih.gov/pubmed/27688432 http://dx.doi.org/10.14814/phy2.12940 |
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