Persistent Calcium Sparklets in Arterial Smooth Muscle

动脉平滑肌中持续存在钙火花

基本信息

批准号：
8308222
负责人：
Luis F Santana
金额：
$ 38.59万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-04-01 至 2017-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8308222
关键词：
Adrenergic Agents Adult Age Agonist Angiotensin II Blood Pressure Blood Vessels Calcineurin Calcium Cartoons Cell Nucleus Cells Cellular Membrane Centers for Disease Control and Prevention (U.S.)Clinical Complex Coupled Coupling Data Development Dihydropyridines Disease Electrophysiology (science)Equilibrium Event Frequencies Functional disorder Funding Gene Expression Genes Genetic Transcription Health Hypertension Knock-in Mouse Knockout Mice Lead Light Location Macromolecular Complexes Methods Modality Modeling Muscle Cells Optics PKC Binding Site Phosphotransferases Physiological Physiological Processes Play Proteins Public Health Regulation Role Sarcolemma Signal Transduction Smooth Muscle Syndrome System Testing Total Internal Reflection Fluorescent Transgenic Organisms Vascular Smooth Muscle Work adrenergic calcineurin phosphatase design dihydropyridine disorder prevention hypertension treatment innovation mutant novel optogenetics patch clamp research study transcription factor transcription factor NF-AT c3 treatment strategy voltage

项目摘要

DESCRIPTION (provided by applicant): The experiments in this application will test the hypothesis that clusters of voltage-gated L-type CaV1.2 channels are capable of undergoing coordinated openings ("coupled gating"), amplifying Ca2+ influx into arterial smooth muscle. A key discovery is that Ca2+ influx via coupled CaV1.2 channels plays a critical role in excitation-contraction (EC) coupling and excitation-transcription (ET) coupling in arterial myocytes. Preliminary data suggest that association of CaV1.2 channels with the anchoring protein AKAP150 is necessary for coupled gating activity in these cells. The significance of these findings is underscored by the observation that the frequency of coupled CaV1.2 gating events increases in arterial smooth muscle during hypertension and that loss of AKAP150 protects against coupled gating and hypertension. The project has two specific aims designed to investigate the mechanisms and physiological implications of these findings. Specific aim 1 is to test the hypothesis that coupled gating of CaV1.2 channels amplifies Ca2+ influx in arterial myocytes. Specific aim 2 is to test the hypothesis that AKAP150 is required for increased coupled CaV1.2 channel activity and the induction of arterial dysfunction during the development of hypertension. The methods that will be used to achieve these aims include patch-clamp electrophysiology, optical clamping, light- and chemically-induced dynamic targeting of kinases to cellular membranes, light-induced activation of adrenergic signaling (i.e., optogenetics), confocal, and TIRF microscopy. Experiments will involve new transgenic, knock in, and knock out mice. This work will generate fundamental information on the mechanisms by which AKAP150 and CaV1.2 channels control of excitability, gene expression, and EC coupling in vascular smooth muscle under physiological and pathological conditions. PUBLIC HEALTH RELEVANCE: Recent data from the Centers for Disease Control and Prevention suggest that, in the USA, nearly 33% of adults ages 20 and older suffer of hypertension, a clinical syndrome characterized by increased vascular tone. However, the mechanisms underlying these pathological changes in vascular function are poorly understood. The experiments proposed in this application have important implications for public health because they investigate the mechanisms by which a new Ca2+ signaling modality regulates the function of vascular smooth muscle under normal conditions and during the development of hypertension. The results of the proposed work may lead to the development of rational strategies for the treatment of hypertension.

描述（由申请人提供）：本申请中的实验将测试电压门控L型CaV1.2通道簇能够协调开放（“耦合门控”）、放大Ca2+流入动脉平滑肌的假设。一个关键的发现是，Ca2+ 通过耦合 CaV1.2 通道的流入在动脉肌细胞的兴奋-收缩 (EC) 耦合和兴奋-转录 (ET) 耦合中发挥着关键作用。初步数据表明，CaV1.2 通道与锚定蛋白 AKAP150 的关联对于这些细胞中的耦合门控活性是必要的。高血压期间动脉平滑肌中耦合 CaV1.2 门控事件的频率增加，并且 AKAP150 的缺失可防止耦合门控和高血压，这一观察结果强调了这些发现的重要性。该项目有两个具体目标，旨在研究这些发现的机制和生理学意义。具体目标 1 是检验 CaV1.2 通道的耦合门控会放大动脉肌细胞中 Ca2+ 流入的假设。具体目标 2 是检验以下假设：在高血压发展过程中，AKAP150 是增加 CaV1.2 通道活性和诱导动脉功能障碍所必需的。用于实现这些目标的方法包括膜片钳电生理学、光钳、光和化学诱导的激酶动态靶向细胞膜、光诱导的肾上腺素信号传导激活（即，光遗传学）、共焦和 TIRF 显微镜。实验将涉及新的转基因、敲入和敲除小鼠。这项工作将产生关于生理和病理条件下 AKAP150 和 CaV1.2 通道控制血管平滑肌兴奋性、基因表达和 EC 耦合的机制的基本信息。公共健康相关性：疾病控制和预防中心的最新数据表明，在美国，近 33% 20 岁及以上的成年人患有高血压，这是一种以血管张力增加为特征的临床综合征。然而，人们对血管功能这些病理变化的机制知之甚少。本申请中提出的实验对公共健康具有重要意义，因为它们研究了新的 Ca2+ 信号传导模式在正常条件下和高血压发展过程中调节血管平滑肌功能的机制。拟议工作的结果可能会导致制定合理的高血压治疗策略。