Regulation of vascular smooth muscle calcium sensitivity

血管平滑肌钙敏感性的调节

• 批准号: 7822205
• 负责人: PAUL H RATZ
• 金额: $ 1.85万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-01 至 2010-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7822205
• 关键词: Agonist; Antibodies; Arachidonic Acids; Biochemical; Blood Vessels; Blood flow; Calcium; Cells; Confocal Microscopy; Contractile Proteins; Contractile System; Contracts; Cyclic AMP-Dependent Protein Kinases; Cyclic Nucleotides; Deterioration; Development; Disease; Docking; Down-Regulation; Enzymes; Failure; Fluorescence; Fura-2; G-Protein-Coupled Receptors; Goals; Hypertension; Knowledge; Laboratories; Link; Maintenance; Measurement; Mediating; Mediator of activation protein; Membrane; Methodology; Modeling; Molecular; Muscle; Muscle Contraction; Myosin Light Chain Kinase; Myosin Light Chains; Oryctolagus cuniculus; Participant; Pathway interactions; Peptide Signal Sequences; Peptides; Phosphoproteins; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphotransferases; Physiological; Play; Procedures; Protein Kinase; Proteins; Regulation; Research; Research Personnel; Role; Series; Shock; Signal Transduction; Signaling Molecule; Site; Small Interfering RNA; Smooth Muscle; Stimulus; System; Techniques; Testing; Therapeutic Agents; Tissues; Up-Regulation; Vascular Smooth Muscle; Vasodilator Agents; Western Blotting; Work; atypical protein kinase C; desensitization; enzyme activity; femoral artery; inhibitor/antagonist; interest; myosin phosphatase; novel; novel therapeutics; programs; protein kinase C kinase; spatiotemporal; tissue culture

DESCRIPTION (provided by applicant): Tissue blood flow is regulated by vascular smooth muscle (VSM) contraction, which in turn, is regulated by changes in the levels of cytosolic free calcium (Ca) and the sensitivity of contractile proteins to Ca. This project will focus on mechanisms regulating rhoA kinase (ROK)-dependent Ca sensitivity. However, what is most novel about this project is that an emphasis will be to understand the cell signaling systems activated by Ca itself that cause ROK-induced Ca sensitization. Regulation of Ca sensitivity is a basic mechanism controlling vascular tone, and dysregulation of Ca sensitivity plays a role in hypertension and the "failure" of smooth muscle to contract in vasodilatory shock. The long-term goal of my laboratory is to investigate subcellular mechanisms regulating VSM Ca sensitivity and tonic force maintenance to provide basic knowledge for the development of novel therapeutic agents to treat selectively vascular contractile disorders. My laboratory has determined that Ca sensitivity can be increased in VSM by a Ca-dependent mechanism. This Ca-dependent Ca sensitization appears to involve activation of ROK and an atypical PKC isotype, PKCzeta, and appears to be dependent on iPLA2 and PI3K activation. The immediate goal of this project is to identify, using physiological, biochemical, pharmacological, cell and molecular, and morphometric methodologies, the molecular mechanisms regulating Ca-dependent Ca sensitivity and tonic force maintenance in a well-characterized arterial contractile system, the KCI-stimulated rabbit FA. The overall goal of this project is to understand regulation of arterial smooth muscle contraction in tissues maintained in as near a physiological state as possible. Whereas this approach has limitations, by applying multiple methodologies to assess spatiotemporal activation of specific signaling molecules proposed to participate in Ca-dependent Ca sensitization, mechanistic conclusions can be drawn regarding cause and effect of discrete steps linking stimulus with contraction in the VSM of intact, functional tissues. The Specific Aim of this study will be to test the hypothesis that ROK and PKCzeta both mediate KCI- induced Ca sensitization of FA, and that iPLA2 and PI3K are required as upstream activators of ROK and PKCzeta.

描述（由申请人提供）：组织血流受血管平滑肌（VSM）收缩调节，而血管平滑肌（VSM）收缩又受胞质游离钙（CA）水平的变化以及收缩蛋白对CA的敏感性的调节。该项目将着重于调节RhoA激酶（ROK）依赖性CA灵敏度的机制。但是，这个项目最新颖的是，重点是了解CA本身激活的细胞信号系统，从而引起ROK诱导的Ca敏感性。 CA灵敏度的调节是控制血管张力的基本机制，CA敏感性失调在高血压和平滑肌在血管舒张性休克中收缩的“失败”起作用。我的实验室的长期目标是研究调节VSM CA敏感性和补体力量维持的亚细胞机制，以为开发新型治疗剂的发展提供基本知识，以治疗有选择的血管收缩障碍。我的实验室确定可以通过CA依赖性机制在VSM中提高CA敏感性。这种依赖于CA的CA敏化似乎涉及ROK的激活和非典型PKC同种型，PKCZETA，并且似乎取决于IPLA2和PI3K激活。该项目的直接目标是使用生理，生化，药理，细胞和分子以及形态计量方法，在良好的特征化的动脉收缩系统中调节CA依赖性CA敏感性和滋补曲肌的分子机制，KCI刺激的兔子FA。该项目的总体目标是了解在尽可能接近生理状态的组织中对动脉平滑肌收缩的调节。尽管这种方法具有局限性，但通过应用多种方法来评估提议参与CA依赖性CA敏化的特定信号传导分子的时空激活，可以得出机械结论，有关离散步骤的原因和效果，将刺激与Intact Intact，功能性组织的刺激联系起来。这项研究的具体目的是检验以下假设：ROK和PKCZETA均介导了FA的CA敏化，并且IPLA2和PI3K是Rok和Pkczeta的上游激活剂所必需的。