Cargando…

Ca(2+) is a Regulator of the WNK/OSR1/NKCC Pathway in a Human Salivary Gland Cell Line

Wnk kinase maintains cell volume, regulating various transporters such as sodium-chloride cotransporter, potassium-chloride cotransporter, and sodium-potassium-chloride cotransporter 1 (NKCC1) through the phosphorylation of oxidative stress responsive kinase 1 (OSR1) and STE20/SPS1-related proline/a...

Descripción completa

Detalles Bibliográficos
Autores principales: Park, Soonhong, Ku, Sang Kyun, Ji, Hye Won, Choi, Jong-Hoon, Shin, Dong Min
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Korean Physiological Society and The Korean Society of Pharmacology 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4422965/
https://www.ncbi.nlm.nih.gov/pubmed/25954130
http://dx.doi.org/10.4196/kjpp.2015.19.3.249
_version_ 1782370129642782720
author Park, Soonhong
Ku, Sang Kyun
Ji, Hye Won
Choi, Jong-Hoon
Shin, Dong Min
author_facet Park, Soonhong
Ku, Sang Kyun
Ji, Hye Won
Choi, Jong-Hoon
Shin, Dong Min
author_sort Park, Soonhong
collection PubMed
description Wnk kinase maintains cell volume, regulating various transporters such as sodium-chloride cotransporter, potassium-chloride cotransporter, and sodium-potassium-chloride cotransporter 1 (NKCC1) through the phosphorylation of oxidative stress responsive kinase 1 (OSR1) and STE20/SPS1-related proline/alanine-rich kinase (SPAK). However, the activating mechanism of Wnk kinase in specific tissues and specific conditions is broadly unclear. In the present study, we used a human salivary gland (HSG) cell line as a model and showed that Ca(2+) may have a role in regulating Wnk kinase in the HSG cell line. Through this study, we found that the HSG cell line expressed molecules participating in the WNK-OSR1-NKCC pathway, such as Wnk1, Wnk4, OSR1, SPAK, and NKCC1. The HSG cell line showed an intracellular Ca(2+) concentration ([Ca(2+)](i)) increase in response to hypotonic stimulation, and the response was synchronized with the phosphorylation of OSR1. Interestingly, when we inhibited the hypotonically induced [Ca(2+)](i) increase with nonspecific Ca(2+) channel blockers such as 2-aminoethoxydiphenyl borate, gadolinium, and lanthanum, the phosphorylated OSR1 level was also diminished. Moreover, a cyclopiazonic acid-induced passive [Ca(2+)](i) elevation was evoked by the phosphorylation of OSR1, and the amount of phosphorylated OSR1 decreased when the cells were treated with BAPTA, a Ca(2+) chelator. Finally, through that process, NKCC1 activity also decreased to maintain the cell volume in the HSG cell line. These results indicate that Ca(2+) may regulate the WNK-OSR1 pathway and NKCC1 activity in the HSG cell line. This is the first demonstration that indicates upstream Ca(2+) regulation of the WNK-OSR1 pathway in intact cells.
format Online
Article
Text
id pubmed-4422965
institution National Center for Biotechnology Information
language English
publishDate 2015
publisher The Korean Physiological Society and The Korean Society of Pharmacology
record_format MEDLINE/PubMed
spelling pubmed-44229652015-05-07 Ca(2+) is a Regulator of the WNK/OSR1/NKCC Pathway in a Human Salivary Gland Cell Line Park, Soonhong Ku, Sang Kyun Ji, Hye Won Choi, Jong-Hoon Shin, Dong Min Korean J Physiol Pharmacol Original Article Wnk kinase maintains cell volume, regulating various transporters such as sodium-chloride cotransporter, potassium-chloride cotransporter, and sodium-potassium-chloride cotransporter 1 (NKCC1) through the phosphorylation of oxidative stress responsive kinase 1 (OSR1) and STE20/SPS1-related proline/alanine-rich kinase (SPAK). However, the activating mechanism of Wnk kinase in specific tissues and specific conditions is broadly unclear. In the present study, we used a human salivary gland (HSG) cell line as a model and showed that Ca(2+) may have a role in regulating Wnk kinase in the HSG cell line. Through this study, we found that the HSG cell line expressed molecules participating in the WNK-OSR1-NKCC pathway, such as Wnk1, Wnk4, OSR1, SPAK, and NKCC1. The HSG cell line showed an intracellular Ca(2+) concentration ([Ca(2+)](i)) increase in response to hypotonic stimulation, and the response was synchronized with the phosphorylation of OSR1. Interestingly, when we inhibited the hypotonically induced [Ca(2+)](i) increase with nonspecific Ca(2+) channel blockers such as 2-aminoethoxydiphenyl borate, gadolinium, and lanthanum, the phosphorylated OSR1 level was also diminished. Moreover, a cyclopiazonic acid-induced passive [Ca(2+)](i) elevation was evoked by the phosphorylation of OSR1, and the amount of phosphorylated OSR1 decreased when the cells were treated with BAPTA, a Ca(2+) chelator. Finally, through that process, NKCC1 activity also decreased to maintain the cell volume in the HSG cell line. These results indicate that Ca(2+) may regulate the WNK-OSR1 pathway and NKCC1 activity in the HSG cell line. This is the first demonstration that indicates upstream Ca(2+) regulation of the WNK-OSR1 pathway in intact cells. The Korean Physiological Society and The Korean Society of Pharmacology 2015-05 2015-04-30 /pmc/articles/PMC4422965/ /pubmed/25954130 http://dx.doi.org/10.4196/kjpp.2015.19.3.249 Text en Copyright © Korean J Physiol Pharmacol and MEDrang Inc. http://creativecommons.org/licenses/by-nc/3.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Article
Park, Soonhong
Ku, Sang Kyun
Ji, Hye Won
Choi, Jong-Hoon
Shin, Dong Min
Ca(2+) is a Regulator of the WNK/OSR1/NKCC Pathway in a Human Salivary Gland Cell Line
title Ca(2+) is a Regulator of the WNK/OSR1/NKCC Pathway in a Human Salivary Gland Cell Line
title_full Ca(2+) is a Regulator of the WNK/OSR1/NKCC Pathway in a Human Salivary Gland Cell Line
title_fullStr Ca(2+) is a Regulator of the WNK/OSR1/NKCC Pathway in a Human Salivary Gland Cell Line
title_full_unstemmed Ca(2+) is a Regulator of the WNK/OSR1/NKCC Pathway in a Human Salivary Gland Cell Line
title_short Ca(2+) is a Regulator of the WNK/OSR1/NKCC Pathway in a Human Salivary Gland Cell Line
title_sort ca(2+) is a regulator of the wnk/osr1/nkcc pathway in a human salivary gland cell line
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4422965/
https://www.ncbi.nlm.nih.gov/pubmed/25954130
http://dx.doi.org/10.4196/kjpp.2015.19.3.249
work_keys_str_mv AT parksoonhong ca2isaregulatorofthewnkosr1nkccpathwayinahumansalivaryglandcellline
AT kusangkyun ca2isaregulatorofthewnkosr1nkccpathwayinahumansalivaryglandcellline
AT jihyewon ca2isaregulatorofthewnkosr1nkccpathwayinahumansalivaryglandcellline
AT choijonghoon ca2isaregulatorofthewnkosr1nkccpathwayinahumansalivaryglandcellline
AT shindongmin ca2isaregulatorofthewnkosr1nkccpathwayinahumansalivaryglandcellline