Mechanisms of cAMP Compartmentation

cAMP 区室的机制

• 批准号: 8733710
• 负责人: ROBERT D HARVEY
• 金额: $ 27.66万
• 依托单位: UNIVERSITY OF NEVADA RENO
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-01 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8733710
• 关键词: Address; Adrenergic Agents; Adrenergic Receptor; Adult; Affect; Behavior; Biological Models; Biosensor; Buffers; Cardiac Myocytes; Cell membrane; Cells; Cellular Structures; Cholesterol; Complex; Computer Simulation; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Cytoplasmic Tail; Cytoskeleton; Data; Diffusion; Disease; Fluorescence Resonance Energy Transfer; Goals; Hormones; Kinetics; Lead; Life; Measurement; Measures; Mechanics; Mediating; Membrane; Metabolic; Methods; Mitochondria; Modeling; Monitor; Muscarinic M2 Receptor; Muscarinics; Muscle Cells; Neurotransmitters; Physiological; Play; Prevention strategy; Problem Solving; Process; Production; Prostaglandin Receptor; Publishing; Rattus; Receptor Activation; Receptor Signaling; Regulation; Role; Sarcoplasmic Reticulum; Second Messenger Systems; Signal Pathway; Signal Transduction; Signaling Molecule; Signaling Protein; Subcellular structure; System; Systems Biology; Testing; Ventricular; adrenergic; base; beta-adrenergic receptor; cell type; innovation; novel strategies; phosphoric diester hydrolase; receptor; receptor-mediated signaling; research study; response; second messenger

DESCRIPTION (provided by applicant): The second messenger cAMP is involved in regulating a variety of responses in virtually every cell in our bodies. One example is the cardiac myocyte, where beta-adrenergic receptors mediate sympathetic effects on electrical, mechanical, and metabolic activity through the production of cAMP. However, there are actually multiple receptors capable of regulating cAMP production in cardiac myocytes, just like many other cells. Even though various receptors may share this common second messenger, they do not all elicit the same functional responses. The common explanation is that cAMP production is compartmentalized, spatially limiting the extent of responses produced by certain receptors. The idea that activation of a receptor does not lead to uniform stimulation or inhibition of cAMP production throughout the cell might seem intuitively obvious, yet it is not fully understood how this is achieved. The common assumption is that phosphodiesterases act as functional barriers to diffusion, creating discrete cAMP signaling domains. We will test the hypothesis that phosphodiesterase activity plays an important role in compartmentation, but not as a barrier to diffusion. We will also test the hypothesis that the cytoskeleton and cholesterol-dependent organization of the plasma membrane play essential roles in this process. The strength of this application lies in the innovative combination of methods that will be used to address these hypotheses. We will employ a systems biology approach that involves the use of multiple fluorescence resonance energy transfer (FRET) based biosensors to monitor cAMP activity in distinct subcellular compartments of live cells, together with quantitative computational modeling of compartmentalized cAMP signaling. The production of cAMP is an important means of eliciting beneficial changes in function of many cell types. However, cAMP-dependent signaling can also produce pathological responses under the right (or wrong) conditions. It has been hypothesized that the difference between the physiological and pathological effects may be a function of appropriate compartmentation of cAMP signaling. Therefore, understanding the contribution of factors that are responsible for coordinating the spatial and temporal distribution of cAMP at the subcellular level could be important for developing new strategies for the prevention or treatment of unfavorable responses associated with many different disease states.

描述（由申请人提供）：第二信使cAMP参与调节我们体内几乎每个细胞的各种反应。一个例子是心肌细胞，其中β-肾上腺素能受体通过产生cAMP介导对电、机械和代谢活动的交感神经效应。然而，实际上有多种受体能够调节心肌细胞中cAMP的产生，就像许多其他细胞一样。尽管不同的受体可能共享这个共同的第二信使，但它们并不都引起相同的功能反应。常见的解释是cAMP的产生是区室化的，在空间上限制了某些受体产生的反应程度。受体的激活不会导致整个细胞中cAMP产生的均匀刺激或抑制的想法似乎直观地显而易见，但尚未完全理解这是如何实现的。通常的假设是磷酸二酯酶作为扩散的功能屏障，产生离散的cAMP信号传导结构域。我们将测试的假设，磷酸二酯酶活性起着重要的作用，在区室化，但不是作为一个障碍扩散。我们还将测试的假设，细胞骨架和细胞膜的胆固醇依赖性组织在这一过程中发挥重要作用。该应用程序的优势在于将用于解决这些假设的方法的创新组合。我们将采用一种系统生物学方法，该方法涉及使用多个基于荧光共振能量转移（FRET）的生物传感器来监测活细胞不同亚细胞区室中的cAMP活性，以及区室化cAMP信号的定量计算建模。cAMP的产生是引发许多细胞类型功能的有益变化的重要手段。然而，cAMP依赖性信号也可以在正确（或错误）的条件下产生病理反应。已经假设生理和病理效应之间的差异可能是cAMP信号传导的适当区室化的功能。因此，了解负责协调cAMP在亚细胞水平上的空间和时间分布的因素的贡献对于开发用于预防或治疗与许多不同疾病状态相关的不利反应的新策略可能是重要的。