| First Author | Garrud TAC | Year | 2024 |
| Journal | Proc Natl Acad Sci U S A | Volume | 121 |
| Issue | 15 | Pages | e2322135121 |
| PubMed ID | 38568964 | Mgi Jnum | J:354616 |
| Mgi Id | MGI:7736141 | Doi | 10.1073/pnas.2322135121 |
| Citation | Garrud TAC, et al. (2024) WNK kinase is a vasoactive chloride sensor in endothelial cells. Proc Natl Acad Sci U S A 121(15):e2322135121 |
| abstractText | 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 Ca(2+) signaling was measured in four dimensions in ECs using a high-speed lightsheet microscope. WNK kinase-dependent activation of TRPV4 channels increased local intracellular Ca(2+) 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. |
