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Regulation of the two-pore domain potassium channel, THIK-1 and THIK-2, by G protein coupled receptors
A member of THIK (two pore domain halothane-inhibited K(+)) channels, THIK-1, was reported as a target of Gi/o-coupled receptors (Gi/o-Rs) in neurons and microglia. We confirmed that in HEK293T cells the THIK-1 channel is activated by Gi/o-Rs and found that Gq-coupled receptors (Gq-Rs) also activate...
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Public Library of Science
2023
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10132538/ https://www.ncbi.nlm.nih.gov/pubmed/37099539 http://dx.doi.org/10.1371/journal.pone.0284962 |
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author | Tateyama, Michihiro Kubo, Yoshihiro |
author_facet | Tateyama, Michihiro Kubo, Yoshihiro |
author_sort | Tateyama, Michihiro |
collection | PubMed |
description | A member of THIK (two pore domain halothane-inhibited K(+)) channels, THIK-1, was reported as a target of Gi/o-coupled receptors (Gi/o-Rs) in neurons and microglia. We confirmed that in HEK293T cells the THIK-1 channel is activated by Gi/o-Rs and found that Gq-coupled receptors (Gq-Rs) also activates the channel. The effects of Gi/o-Rs and Gq-Rs were inhibited by the Gi/o inhibitor pertussis toxin and phospholipase C (PLC) inhibitor, respectively. The effects of Gi/o-Rs were attenuated when consensus Gβγ binding motif at the C-tail of the THIK-1 channel was mutated, suggesting that Gβγ serves as a THIK-1 channel activator upon the stimulation of Gi/o-Rs. As to the effects of Gq-Rs on the THIK-1 channel, a protein kinase C inhibitor and calcium chelators failed to inhibit the effect of a Gq coupled muscarinic M1R. Neither the hydrolysis of phosphatidyl inositol bisphosphate induced by voltage sensitive phosphatase nor the application of a diacylglycerol analogue, OAG, increased the channel current. The mediator of Gq-dependent activation of the THIK-1 channel remained unsolved. The effects of Gi/o- and Gq-Rs on the THIK-2 channel were also investigated, by using a THIK-2 mutant channel whose N-terminal domain is deleted to improve the surface membrane expression. We observed that Gi/o- and Gq-Rs activate the mutated THIK-2 channel, similarly to the THIK-1 channel. Interestingly, heterodimeric channels of THIK-1 and THIK-2 responded to Gi/o-R and Gq-R stimulation. Taken together, Gi/o- or Gq-Rs activates the THIK-1 and THIK-2 channels in a Gβγ or PLC dependent manner, respectively. |
format | Online Article Text |
id | pubmed-10132538 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-101325382023-04-27 Regulation of the two-pore domain potassium channel, THIK-1 and THIK-2, by G protein coupled receptors Tateyama, Michihiro Kubo, Yoshihiro PLoS One Research Article A member of THIK (two pore domain halothane-inhibited K(+)) channels, THIK-1, was reported as a target of Gi/o-coupled receptors (Gi/o-Rs) in neurons and microglia. We confirmed that in HEK293T cells the THIK-1 channel is activated by Gi/o-Rs and found that Gq-coupled receptors (Gq-Rs) also activates the channel. The effects of Gi/o-Rs and Gq-Rs were inhibited by the Gi/o inhibitor pertussis toxin and phospholipase C (PLC) inhibitor, respectively. The effects of Gi/o-Rs were attenuated when consensus Gβγ binding motif at the C-tail of the THIK-1 channel was mutated, suggesting that Gβγ serves as a THIK-1 channel activator upon the stimulation of Gi/o-Rs. As to the effects of Gq-Rs on the THIK-1 channel, a protein kinase C inhibitor and calcium chelators failed to inhibit the effect of a Gq coupled muscarinic M1R. Neither the hydrolysis of phosphatidyl inositol bisphosphate induced by voltage sensitive phosphatase nor the application of a diacylglycerol analogue, OAG, increased the channel current. The mediator of Gq-dependent activation of the THIK-1 channel remained unsolved. The effects of Gi/o- and Gq-Rs on the THIK-2 channel were also investigated, by using a THIK-2 mutant channel whose N-terminal domain is deleted to improve the surface membrane expression. We observed that Gi/o- and Gq-Rs activate the mutated THIK-2 channel, similarly to the THIK-1 channel. Interestingly, heterodimeric channels of THIK-1 and THIK-2 responded to Gi/o-R and Gq-R stimulation. Taken together, Gi/o- or Gq-Rs activates the THIK-1 and THIK-2 channels in a Gβγ or PLC dependent manner, respectively. Public Library of Science 2023-04-26 /pmc/articles/PMC10132538/ /pubmed/37099539 http://dx.doi.org/10.1371/journal.pone.0284962 Text en © 2023 Tateyama, Kubo https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Tateyama, Michihiro Kubo, Yoshihiro Regulation of the two-pore domain potassium channel, THIK-1 and THIK-2, by G protein coupled receptors |
title | Regulation of the two-pore domain potassium channel, THIK-1 and THIK-2, by G protein coupled receptors |
title_full | Regulation of the two-pore domain potassium channel, THIK-1 and THIK-2, by G protein coupled receptors |
title_fullStr | Regulation of the two-pore domain potassium channel, THIK-1 and THIK-2, by G protein coupled receptors |
title_full_unstemmed | Regulation of the two-pore domain potassium channel, THIK-1 and THIK-2, by G protein coupled receptors |
title_short | Regulation of the two-pore domain potassium channel, THIK-1 and THIK-2, by G protein coupled receptors |
title_sort | regulation of the two-pore domain potassium channel, thik-1 and thik-2, by g protein coupled receptors |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10132538/ https://www.ncbi.nlm.nih.gov/pubmed/37099539 http://dx.doi.org/10.1371/journal.pone.0284962 |
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