Table_2_Stimulation of the Epithelial Na+ Channel in Renal Principal Cells by Gs-Coupled Designer Receptors Exclusively Activated by Designer Drugs.DOCX
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.
History
References
- https://doi.org//10.1038/s41598-021-94118-3
- https://doi.org//10.1007/978-1-4939-9121-1_21
- https://doi.org//10.1128/MCB.00584-16
- https://doi.org//10.1152/ajprenal.00469.2017
- https://doi.org//10.1152/ajpcell.00301.2011
- https://doi.org//10.1152/ajprenal.00371.2009
- https://doi.org//10.1038/361467a0
- https://doi.org//10.1038/367463a0
- https://doi.org//10.1681/ASN.2006080902
- https://doi.org//10.1016/j.gene.2015.12.061
- https://doi.org//10.1152/ajpcell.1992.263.4.C825
- https://doi.org//10.1371/journal.pbio.1000412
- https://doi.org//10.1152/ajprenal.00505.2013
- https://doi.org//10.1073/pnas.1201978109
- https://doi.org//10.1007/978-1-62703-351-0_27
- https://doi.org//10.1152/ajprenal.00246.2014
- https://doi.org//10.1152/ajprenal.00612.2018
- https://doi.org//10.1681/ASN.2008010021
- https://doi.org//10.1172/JCI112433
- https://doi.org//10.1681/ASN.2010040409
- https://doi.org//10.1681/ASN.2012050449
- https://doi.org//10.1152/ajprenal.00397.2011
- https://doi.org//10.1146/annurev.physiol.64.082101.143243
- https://doi.org//10.1172/JCI7869
- https://doi.org//10.1074/jbc.M407858200
- https://doi.org//10.1529/biophysj.104.056804
- https://doi.org//10.1385/1-59745-095-2%3A3
- https://doi.org//10.1038/ki.2010.276
- https://doi.org//10.3389/fphys.2012.00304
- https://doi.org//10.1002/cphy.c100051
- https://doi.org//10.1152/ajprenal.00109.2009
- https://doi.org//10.1159/000020587
- https://doi.org//10.1016/j.coph.2014.12.012