WNK kinase is a vasoactive chloride sensor in endothelial cells

• 成果类型: Article
• 署名作者: Garrud, Tessa A. C.; Bell, Briar; Mata-Daboin, Alejandro; Peixoto-Neves, Dieniffer; Collier, Daniel M.; Cordero-Morales, Julio F.; Jaggar, Jonathan H.
• 署名单位: University of Tennessee System; University of Tennessee Health Science Center; University of Texas System; University of Texas Health Science Center Houston; University of Tennessee System; University of Tennessee Health Science Center
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-9811
• DOI: 10.1073/pnas.2322135121
• 发表日期: 2024-04-09
• 关键词: spak/osr1 kinases; protein-kinases; cl; expression; channels; tmem16a; phosphorylation; ca2+; homeostasis; activation

摘要

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 late EC function to alter arterial contractility is unclear. Here, we tested the hypothesis 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 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.