Faculty, Staff and Student Publications
Language
English
Publication Date
4-9-2024
Journal
Proceedings of the National Academy of Sciences of the United States of America
DOI
10.1073/pnas.2322135121
PMID
38568964
PMCID
PMC11009681
PubMedCentral® Posted Date
4-3-2024
PubMedCentral® Full Text Version
Post-print
Abstract
Endothelial cells (ECs) line the wall of blood vessels and regulate arterial contractility to tune regional organ blood flow and systemic pressure. Chloride (Cl-) is the most abundant anion in ECs and the Cl- sensitive With-No-Lysine (WNK) kinase is expressed in this cell type. Whether intracellular Cl- signaling and WNK kinase regulate EC function to alter arterial contractility is unclear. Here, we tested the hypothesis that intracellular Cl- signaling in ECs regulates arterial contractility and examined the signaling mechanisms involved, including the participation of WNK kinase. Our data obtained using two-photon microscopy and cell-specific inducible knockout mice indicated that acetylcholine, a prototypical vasodilator, stimulated a rapid reduction in intracellular Cl- concentration ([Cl-]i) due to the activation of TMEM16A, a Cl- channel, in ECs of resistance-size arteries. TMEM16A channel-mediated Cl- signaling activated WNK kinase, which phosphorylated its substrate proteins SPAK and OSR1 in ECs. OSR1 potentiated transient receptor potential vanilloid 4 (TRPV4) currents in a kinase-dependent manner and required a conserved binding motif located in the channel C terminus. Intracellular Ca2+ signaling was measured in four dimensions in ECs using a high-speed lightsheet microscope. WNK kinase-dependent activation of TRPV4 channels increased local intracellular Ca2+ signaling in ECs and produced vasodilation. In summary, we show that TMEM16A channel activation reduces [Cl-]i, which activates WNK kinase in ECs. WNK kinase phosphorylates OSR1 which then stimulates TRPV4 channels to produce vasodilation. Thus, TMEM16A channels regulate intracellular Cl- signaling and WNK kinase activity in ECs to control arterial contractility.
Keywords
Mice, Animals, Protein Serine-Threonine Kinases, Chlorides, Endothelial Cells, TRPV Cation Channels, Signal Transduction, TMEM16A, WNK kinase, TRPV4, endothelial cell, vasodilation
Published Open-Access
yes
Recommended Citation
Garrud, Tessa A C; Bell, Briar; Mata-Daboin, Alejandro; et al., "Wnk Kinase Is a Vasoactive Chloride Sensor in Endothelial Cells" (2024). Faculty, Staff and Student Publications. 2844.
https://digitalcommons.library.tmc.edu/uthmed_docs/2844