TY - JOUR

T1 - Rapid Aldosterone-Mediated Signaling in the DCT Increases Activity of the Thiazide-Sensitive NaCl Cotransporter

AU - Cheng, Lei

AU - Poulsen, Soren Brandt

AU - Wu, Qi

AU - Esteva-Font, Cristina

AU - Olesen, Emma T. B.

AU - Peng, Li

AU - Olde, Bjorn

AU - Leeb-Lundberg, L. M. Fredrik

AU - Pisitkun, Trairak

AU - Rieg, Timo

AU - Dimke, Henrik

AU - Fenton, Robert A.

PY - 2019

Y1 - 2019

N2 - Background The NaCl cotransporter NCC in the kidney distal convoluted tubule (DCT) regulates urinary NaCl excretion and BP. Aldosterone increases NaCl reabsorption via NCC over the long-term by altering gene expression. But the acute effects of aldosterone in the DCT are less well understood.Methods Proteomics, bioinformatics, and cell biology approaches were combined with animal models and gene-targeted mice.Results Aldosterone significantly increases NCC activity within minutes in vivo or ex vivo. These effects were independent of transcription and translation, but were absent in the presence of high potassium. In vitro, aldosterone rapidly increased intracellular cAMP and inositol phosphate accumulation, and altered phosphorylation of various kinases/kinase substrates within the MAPK/ERK, PI3K/AKT, and cAMP/PKA pathways. Inhibiting GPR30, a membrane-associated receptor, limited aldosterone's effects on NCC activity ex vivo, and NCC phosphorylation was reduced in GPR30 knockout mice. Phosphoproteomics, network analysis, and in vitro studies determined that aldosterone activates EGFR-dependent signaling. The EGFR immunolocalized to the DCT and EGFR tyrosine kinase inhibition decreased NCC activity ex vivo and in vivo.Conclusions Aldosterone acutely activates NCC to modulate renal NaCl excretion.

AB - Background The NaCl cotransporter NCC in the kidney distal convoluted tubule (DCT) regulates urinary NaCl excretion and BP. Aldosterone increases NaCl reabsorption via NCC over the long-term by altering gene expression. But the acute effects of aldosterone in the DCT are less well understood.Methods Proteomics, bioinformatics, and cell biology approaches were combined with animal models and gene-targeted mice.Results Aldosterone significantly increases NCC activity within minutes in vivo or ex vivo. These effects were independent of transcription and translation, but were absent in the presence of high potassium. In vitro, aldosterone rapidly increased intracellular cAMP and inositol phosphate accumulation, and altered phosphorylation of various kinases/kinase substrates within the MAPK/ERK, PI3K/AKT, and cAMP/PKA pathways. Inhibiting GPR30, a membrane-associated receptor, limited aldosterone's effects on NCC activity ex vivo, and NCC phosphorylation was reduced in GPR30 knockout mice. Phosphoproteomics, network analysis, and in vitro studies determined that aldosterone activates EGFR-dependent signaling. The EGFR immunolocalized to the DCT and EGFR tyrosine kinase inhibition decreased NCC activity ex vivo and in vivo.Conclusions Aldosterone acutely activates NCC to modulate renal NaCl excretion.

U2 - 10.1681/ASN.2018101025

DO - 10.1681/ASN.2018101025

M3 - Journal article

C2 - 31253651

VL - 30

SP - 1453

EP - 1469

JO - Journal of the American Society of Nephrology : JASN

JF - Journal of the American Society of Nephrology : JASN

SN - 1046-6673

IS - 8

ER -