We recently reported that the inward-rectifier Kir2.1 channel in brain capillary endothelial cells (cECs) plays a major role in neurovascular coupling (NVC) by mediating a neuronal activity-dependent, propagating vasodilatory (hyperpolarizing) signal. We further demonstrated that Kir2.1 activity is suppressed by depletion of plasma membrane phosphatidylinositol 4,5-bisphosphate (PIP₂). Whether cECs express depolarizing channels that intersect with Kir2.1-mediated signaling remains unknown. Here, we report that Ca²⁺/Na⁺-permeable TRPV4 (transient receptor potential vanilloid 4) channels are expressed in cECs and are tonically inhibited by PIP₂. We further demonstrate that depletion of PIP₂ by agonists, including putative NVC mediators, that promote PIP₂ hydrolysis by signaling through G_q-protein-coupled receptors (G_qPCRs) caused simultaneous disinhibition of TRPV4 channels and suppression of Kir2.1 channels. These findings collectively support the concept that G_qPCR activation functions as a molecular switch to favor capillary TRPV4 activity over Kir2.1 signaling, an observation with potentially profound significance for the control of cerebral blood flow.

Mechanical forces influence different cell types in our bodies. Among the earliest forces experienced in mammals is blood movement in the vascular system. Blood flow starts at the embryonic stage and ceases when the heart stops. Blood flow exposes endothelial cells (ECs) that line all blood vessels to hemodynamic forces. ECs detect these mechanical forces (mechanosensing) through mechanosensors, thus triggering physiological responses such as changes in vascular diameter. In this review, we focus on endothelial mechanosensing and on how different ion channels, receptors, and membrane structures detect forces and mediate intricate mechanotransduction responses. We further highlight that these responses often reflect collaborative efforts involving several mechanosensors and mechanotransducers. We close with a consideration of current knowledge regarding the dysregulation of endothelial mechanosensing during disease. Because hemodynamic disruptions are hallmarks of cardiovascular disease, studying endothelial mechanosensing holds great promise for advancing our understanding of vascular physiology and pathophysiology.

Capillaries are equipped to sense neurovascular coupling agents released onto the outer wall of a capillary, translating these external signals into electrical/Ca ²⁺ changes that play a crucial role in blood flow regulation and ensuring that neuronal demands are met. However, control mechanisms attributable to forces imposed onto the lumen are less clear. Here, we show that Piezo1 channels act as mechanosensors in central nervous system capillaries. Electrophysiological analyses confirmed expression and function of Piezo1 channels in brain cortical and retinal capillaries. Activation of Piezo1 channels evoked currents that were sensitive to endothelial cell–specific Piezo1 deletion. Using genetically encoded Ca ²⁺ indicator mice and an ex vivo pressurized retina preparation, we found that activation of Piezo1 channels by mechanical forces triggered Ca ²⁺ signals in capillary endothelial cells. Collectively, these findings indicate that Piezo1 channels are capillary mechanosensors that initiate crucial Ca ²⁺ signals and could, therefore, have a profound impact on central nervous system blood flow control.

Brain capillary calcium signaling controls blood flow.

Significance: Years before the emergence of infarctions or significant cognitive decline, patients with cerebral small vessel disease (SVD) show a deterioration in the ability of the brain to augment blood flow locally in response to increases in neuronal activity (functional hyperemia). Using a well-characterized genetic mouse model of a hereditary form of SVD, we determined the molecular defect at play in capillary endothelial cells. We found that SVD is associated with reduced synthesis of the phospholipid PIP ₂ , which prevents the Kir2.1 channel-initiated capillary-to-arteriole electrical signaling that supports vasodilatory responses during functional hyperemia. We further show that systemic injection of exogenous PIP ₂ is sufficient to rescue this deficit in SVD mice, restoring adequate cerebral blood flow in response to neuronal activation.

Significance: Capillaries, the smallest blood vessels, mediate the on-demand delivery of oxygen and nutrients required to support the function of active cells throughout the brain. But how blood flow is directed to cells in active brain regions to satisfy their energy needs is poorly understood. We demonstrate that the plasma membrane phospholipid, PIP ₂ , is fundamental to sustaining the activity of inwardly rectifying potassium channels—the molecular feature that allows capillary endothelial cells to sense ongoing neuronal activity and trigger an increase in local blood flow. We further show that chemical factors released in the brain, including those associated with neuronal activity, cause changes in the levels of PIP ₂ , thereby altering endothelial potassium channel signaling and controlling cerebral blood flow.