Cargando…
Stimulation of the Epithelial Na(+) Channel in Renal Principal Cells by Gs-Coupled Designer Receptors Exclusively Activated by Designer Drugs
The activity of the Epithelial Na(+) Channel (ENaC) in renal principal cells (PC) fine-tunes sodium excretion and consequently, affects blood pressure. The Gs-adenylyl cyclase-cAMP signal transduction pathway is believed to play a central role in the normal control of ENaC activity in PCs. The curre...
Autores principales: | , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8425396/ https://www.ncbi.nlm.nih.gov/pubmed/34512393 http://dx.doi.org/10.3389/fphys.2021.725782 |
_version_ | 1783749843739475968 |
---|---|
author | Soares, Antonio G. Contreras, Jorge Archer, Crystal R. Mironova, Elena Berdeaux, Rebecca Stockand, James D. Abd El-Aziz, Tarek Mohamed |
author_facet | Soares, Antonio G. Contreras, Jorge Archer, Crystal R. Mironova, Elena Berdeaux, Rebecca Stockand, James D. Abd El-Aziz, Tarek Mohamed |
author_sort | Soares, Antonio G. |
collection | PubMed |
description | The activity of the Epithelial Na(+) Channel (ENaC) in renal principal cells (PC) fine-tunes sodium excretion and consequently, affects blood pressure. The Gs-adenylyl cyclase-cAMP signal transduction pathway is believed to play a central role in the normal control of ENaC activity in PCs. The current study quantifies the importance of this signaling pathway to the regulation of ENaC activity in vivo using a knock-in mouse that has conditional expression of Gs-DREADD (designer receptors exclusively activated by designer drugs; GsD) in renal PCs. The GsD mouse also contains a cAMP response element-luciferase reporter transgene for non-invasive bioluminescence monitoring of cAMP signaling. Clozapine N-oxide (CNO) was used to selectively and temporally stimulate GsD. Treatment with CNO significantly increased luciferase bioluminescence in the kidneys of PC-specific GsD but not control mice. CNO also significantly increased the activity of ENaC in principal cells in PC-specific GsD mice compared to untreated knock-in mice and CNO treated littermate controls. The cell permeable cAMP analog, 8-(4-chlorophenylthio)adenosine 3′,5′-cyclic monophosphate, significantly increased the activity and expression in the plasma membrane of recombinant ENaC expressed in CHO and COS-7 cells, respectively. Treatment of PC-specific GsD mice with CNO rapidly and significantly decreased urinary Na(+) excretion compared to untreated PC-specific GsD mice and treated littermate controls. This decrease in Na(+) excretion in response to CNO in PC-specific GsD mice was similar in magnitude and timing as that induced by the selective vasopressin receptor 2 agonist, desmopressin, in wild type mice. These findings demonstrate for the first time that targeted activation of Gs signaling exclusively in PCs is sufficient to increase ENaC activity and decrease dependent urinary Na(+) excretion in live animals. |
format | Online Article Text |
id | pubmed-8425396 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-84253962021-09-09 Stimulation of the Epithelial Na(+) Channel in Renal Principal Cells by Gs-Coupled Designer Receptors Exclusively Activated by Designer Drugs Soares, Antonio G. Contreras, Jorge Archer, Crystal R. Mironova, Elena Berdeaux, Rebecca Stockand, James D. Abd El-Aziz, Tarek Mohamed Front Physiol Physiology The activity of the Epithelial Na(+) Channel (ENaC) in renal principal cells (PC) fine-tunes sodium excretion and consequently, affects blood pressure. The Gs-adenylyl cyclase-cAMP signal transduction pathway is believed to play a central role in the normal control of ENaC activity in PCs. The current study quantifies the importance of this signaling pathway to the regulation of ENaC activity in vivo using a knock-in mouse that has conditional expression of Gs-DREADD (designer receptors exclusively activated by designer drugs; GsD) in renal PCs. The GsD mouse also contains a cAMP response element-luciferase reporter transgene for non-invasive bioluminescence monitoring of cAMP signaling. Clozapine N-oxide (CNO) was used to selectively and temporally stimulate GsD. Treatment with CNO significantly increased luciferase bioluminescence in the kidneys of PC-specific GsD but not control mice. CNO also significantly increased the activity of ENaC in principal cells in PC-specific GsD mice compared to untreated knock-in mice and CNO treated littermate controls. The cell permeable cAMP analog, 8-(4-chlorophenylthio)adenosine 3′,5′-cyclic monophosphate, significantly increased the activity and expression in the plasma membrane of recombinant ENaC expressed in CHO and COS-7 cells, respectively. Treatment of PC-specific GsD mice with CNO rapidly and significantly decreased urinary Na(+) excretion compared to untreated PC-specific GsD mice and treated littermate controls. This decrease in Na(+) excretion in response to CNO in PC-specific GsD mice was similar in magnitude and timing as that induced by the selective vasopressin receptor 2 agonist, desmopressin, in wild type mice. These findings demonstrate for the first time that targeted activation of Gs signaling exclusively in PCs is sufficient to increase ENaC activity and decrease dependent urinary Na(+) excretion in live animals. Frontiers Media S.A. 2021-08-25 /pmc/articles/PMC8425396/ /pubmed/34512393 http://dx.doi.org/10.3389/fphys.2021.725782 Text en Copyright © 2021 Soares, Contreras, Archer, Mironova, Berdeaux, Stockand and Abd El-Aziz. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Physiology Soares, Antonio G. Contreras, Jorge Archer, Crystal R. Mironova, Elena Berdeaux, Rebecca Stockand, James D. Abd El-Aziz, Tarek Mohamed Stimulation of the Epithelial Na(+) Channel in Renal Principal Cells by Gs-Coupled Designer Receptors Exclusively Activated by Designer Drugs |
title | Stimulation of the Epithelial Na(+) Channel in Renal Principal Cells by Gs-Coupled Designer Receptors Exclusively Activated by Designer Drugs |
title_full | Stimulation of the Epithelial Na(+) Channel in Renal Principal Cells by Gs-Coupled Designer Receptors Exclusively Activated by Designer Drugs |
title_fullStr | Stimulation of the Epithelial Na(+) Channel in Renal Principal Cells by Gs-Coupled Designer Receptors Exclusively Activated by Designer Drugs |
title_full_unstemmed | Stimulation of the Epithelial Na(+) Channel in Renal Principal Cells by Gs-Coupled Designer Receptors Exclusively Activated by Designer Drugs |
title_short | Stimulation of the Epithelial Na(+) Channel in Renal Principal Cells by Gs-Coupled Designer Receptors Exclusively Activated by Designer Drugs |
title_sort | stimulation of the epithelial na(+) channel in renal principal cells by gs-coupled designer receptors exclusively activated by designer drugs |
topic | Physiology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8425396/ https://www.ncbi.nlm.nih.gov/pubmed/34512393 http://dx.doi.org/10.3389/fphys.2021.725782 |
work_keys_str_mv | AT soaresantoniog stimulationoftheepithelialnachannelinrenalprincipalcellsbygscoupleddesignerreceptorsexclusivelyactivatedbydesignerdrugs AT contrerasjorge stimulationoftheepithelialnachannelinrenalprincipalcellsbygscoupleddesignerreceptorsexclusivelyactivatedbydesignerdrugs AT archercrystalr stimulationoftheepithelialnachannelinrenalprincipalcellsbygscoupleddesignerreceptorsexclusivelyactivatedbydesignerdrugs AT mironovaelena stimulationoftheepithelialnachannelinrenalprincipalcellsbygscoupleddesignerreceptorsexclusivelyactivatedbydesignerdrugs AT berdeauxrebecca stimulationoftheepithelialnachannelinrenalprincipalcellsbygscoupleddesignerreceptorsexclusivelyactivatedbydesignerdrugs AT stockandjamesd stimulationoftheepithelialnachannelinrenalprincipalcellsbygscoupleddesignerreceptorsexclusivelyactivatedbydesignerdrugs AT abdelaziztarekmohamed stimulationoftheepithelialnachannelinrenalprincipalcellsbygscoupleddesignerreceptorsexclusivelyactivatedbydesignerdrugs |