The Role of Bioactive Lipids in Transient Receptor Potential Channels Gating

血压调节依赖于膜蛋白传递物理刺激变化的能力 (例如，血流动力)转化为电信号。瞬时受体电位香草素4(TRPV4)是一种非特异性PU。 机械敏感钙通道在血管内皮细胞、平滑肌细胞和血管周围的表达 感觉神经元。尽管TRPV4与内皮和血管周围神经依赖有关 血管松弛，其确切的门控机制仍然难以捉摸。机械刺激后激活TRPV4的机制有三种：1)磷脂酶A2(PLA2)下游依赖于omega(W)-6花生四烯酸(AA)及其代谢物环氧二十碳三烯酸(EETs)的形成；2)嘌呤能P2Y2受体下游由三磷酸腺苷释放介导的激活；3)膜拉伸直接激活。我们的长期目标是描述离子通道解码外源性和内源性刺激以调节细胞功能的机制。在这项提案中，总体目标是建立TRPV4激活的分子基础。中心假设是TRPV4的激活是 通过脂质重塑来调节膜的机械性能。建议的理由是 研究计划是，一旦阐明了TRPV4激活的确切机制，就有可能 确定以TRPV4为靶点的控制全身血压的策略。这一假设将通过以下方式进行检验 追求三个具体目标：1)确定w-6和w-3脂肪酸对线虫TRPV4活性的影响；2)验证w-3脂肪酸衍生物增强血管细胞TRPV4活性的假设；以及3) 确定膜机械性能的变化如何调节TRPV4门控。我们将利用遗传、行为、功能、生化和生物物理方法来揭示脂肪酸及其代谢物对TRPV4功能的贡献。这项研究计划具有创新性，因为它将决定 W-3和w-6脂肪酸及其二十烷基衍生物对trpv4门控的个体贡献。这项拟议的研究具有重要意义，因为它有望在针对TRPV4的研究中具有广泛的翻译重要性 调节血管和神经功能。

Blood pressure regulation relies on the ability of membrane proteins to transduce variations in physical stimuli (e.g., hemodynamic forces) into electrical signals. The transient receptor potential vanilloid 4 (TRPV4) is a pu- tative mechanosensitive Ca2+ channel expressed in endothelial and smooth muscle cells and in perivascular sensory neurons. Although TRPV4 has been implicated in endothelium- and perivascular nerve-dependent vasorelaxation, its precise gating mechanism remains elusive. Three mechanisms have been proposed to activate TRPV4 after mechanical stimulation: 1) downstream of the phospholipase A2 (PLA2)-dependent formation of omega (w)-6 arachidonic acid (AA) and its metabolites, epoxyeicosatrienoic acids (EETs); 2) downstream of purinergic P2Y2 receptor activation, mediated by adenosine triphosphate release; and 3) direct activation by membrane stretch. Our long-term goal is to delineate the mechanisms by which ion channels decode exogenous and endogenous stimuli to regulate cellular function. In this proposal, the overall objective is to establish the molecular basis underlying TRPV4 activation. The central hypothesis is that TRPV4 activation is regulated by the mechanical properties of the membrane via lipid remodeling. The rationale for the proposed research plan is that once the precise mechanism of TRPV4 activation has been elucidated, it will be possible to define strategies that target TRPV4 to control systemic blood pressure. The hypothesis will be tested by pursuing three Specific Aims: 1) Determine the effect of w-6 and w-3 fatty acids on TRPV4 activity in C. elegans; 2) Test the hypothesis that w-3 fatty acid derivatives enhance TRPV4 activity in vascular cells; and 3) Determine how changes in the mechanical properties of the membrane regulates TRPV4 gating. We will leverage genetic, behavioral, functional, biochemical, and biophysical approaches to uncover the contribution of fatty acids and their metabolites to TRPV4 function. The research plan is innovative because it will determine the individual contribution of w-3 and w-6 fatty acids and their eicosanoid derivatives to TRPV4 gating. The proposed research is significant because it is expected to have broad translational importance in targeting TRPV4 to regulate vascular and neuronal function.