Transient receptor potential vanilloid 4 channels (TRPV4) are pivotal regulators of vascular homeostasis. Altered TRPV4 signaling has recently been implicated in various cardiovascular diseases, including hypertension and atherosclerosis. These versatile nonselective cation channels increase endothelial Ca²⁺ influx in response to various stimuli including shear stress and G protein-coupled receptor (GPCR) activation. Recent findings suggest TRPV4 channels produce localized Ca²⁺ transients at the endothelial cell plasma membrane that may allow targeted effector recruitment and promote large-scale Ca²⁺ events via release from internal stores (endoplasmic reticulum). However, the specific impact of TRPV4 channels on Ca²⁺ signaling in the intact arterial intima remains unknown. In the current study, we employ an endothelium-specific TRPV4 knockout mouse model (ecTRPV4^−/−) to identify and characterize TRPV4-dependent endothelial Ca²⁺ dynamics. We find that carotid arteries from both ecTRPV4^−/− and WT mice exhibit a range of basal and acetylcholine (ACh)-induced Ca²⁺ dynamics, similar in net frequency. Analysis of discrete Ca²⁺ event parameters (amplitude, duration, and spread) and event composite values reveals that while ecTRPV4^−/− artery endothelium predominantly produces large Ca²⁺ events comparable to and in excess of those produced by WT endothelium, they are deficient in a particular population of small events, under both basal and ACh-stimulated conditions. These findings support the concept that TRPV4 channels are responsible for generating a distinct population of focal Ca²⁺ transients in the intact arterial endothelium, likely underlying their essential role in vascular homeostasis.

We hypothesized that transgenic mice overexpressing thep22phoxsubunit of the NADPH oxidase selectively in smooth muscle (Tgp22smc) would exhibit an exacerbated response to transluminal carotid injury compared to wild-type mice. To examine the role of reactive oxygen species (ROS) as a mediator of vascular injury, the injury response was quantified by measuring wall thickness (WT) and cross-sectional wall area (CSWA) of the injured and noninjured arteries in bothTgp22smcand wild-type animals at days 3, 7, and 14 after injury. Akt, p38 MAPK, and Src activation were evaluated at the same time points using Western blotting. WT and CSWA following injury were significantly greater inTgp22smcmice at both 7 and 14 days after injury while noninjured contralateral carotids were similar between groups. Apocynin treatment attenuated the injury response in both groups and rendered the response similar betweenTgp22smcmice and wild-type mice. Following injury, carotid arteries fromTgp22smcmice demonstrated elevated activation of Akt at day 3, while p38 MAPK and Src activation was elevated at day 7 compared to wild-type mice. Both increased activation and temporal regulation of these signaling pathways may contribute to enhanced vascular growth in response to injury in this transgenic model of elevated vascular ROS.