NOS1/NOS3 Functional Effects on Cardiac Myocyte Function

NOS1/NOS3 对心肌细胞功能的影响

• 批准号: 7868057
• 负责人: MARK T ZIOLO
• 金额: $ 36.41万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7868057
• 关键词: Action Potentials; Adrenergic Agents; Arts; Biochemical; Biological Assay; Ca(2+)-Transporting ATPase; Cardiac; Cardiac Myocytes; Cells; Coupling; Cyclic GMP; Cyclic GMP-Dependent Protein Kinases; Data; Disease; Electrophysiology (science); Enzymes; Frequencies; Functional disorder; Heart; Heart Diseases; Heart failure; Human; Knock-out; Knockout Mice; Measures; Muscle Cells; NOS1 gene; NOS3 gene; Neurons; Nitric Oxide; Nitric Oxide Synthase; Nitric Oxide Synthase Type I; Optical Methods; Oryctolagus cuniculus; Pathway interactions; Peroxonitrite; Phosphoric Monoester Hydrolases; Phosphorylation; Physiological; Play; Production; Protein Isoforms; Proteins; Public Health; Regulation; Research Personnel; Role; Sarcoplasmic Reticulum; Signal Pathway; Signal Transduction; Western World; adrenergic; design; drug development; human NOS3 protein; improved; inhibitor/antagonist; mortality; nitration; phospholamban; programs; research study; response; Action Potentials; Adrenergic Agents; Arts; Biochemical; Biological Assay; Ca(2+)-Transporting ATPase; Cardiac; Cardiac Myocytes; Cells; Coupling; Cyclic GMP; Cyclic GMP-Dependent Protein Kinases; Data; Disease; Electrophysiology (science); Enzymes; Frequencies; Functional disorder; Heart; Heart Diseases; Heart failure; Human; Knock-out; Knockout Mice; Measures; Muscle Cells; NOS1 gene; NOS3 gene; Neurons; Nitric Oxide; Nitric Oxide Synthase; Nitric Oxide Synthase Type I; Optical Methods; Oryctolagus cuniculus; Pathway interactions; Peroxonitrite; Phosphoric Monoester Hydrolases; Phosphorylation; Physiological; Play; Production; Protein Isoforms; Proteins; Public Health; Regulation; Research Personnel; Role; Sarcoplasmic Reticulum; Signal Pathway; Signal Transduction; Western World; adrenergic; design; drug development; human NOS3 protein; improved; inhibitor/antagonist; mortality; nitration; phospholamban; programs; research study; response

DESCRIPTION (provided by applicant): Cardiac myocytes constitutively express enzymes termed neuronal nitric oxide synthase (nNOS, NOS1) and endothelial nitric oxide synthase (eNOS, NOS3). Nitric oxide (NO) produced via NOS1 and NOS3 is an important regulator of cardiac contractility in the normal, healthy heart. NOS1 and NOS3 also play a key role in the remodeling and cardiac dysfunction of diseased hearts. NOS1 has been found to be essential for a positive force-frequency response and to potentiate the beta-adrenergic response. This is controversial, as other studies have found somewhat dissimilar results. The effects of NOS3 are better understood, in that NOS3 limits the beta-adrenergic response. However, definitive mechanisms, end targets and signaling pathway(s) for NOS1 and NOS3 are not known. Thus, the objective of this proposal is to critically examine how NOS1 and NOS3 regulate cardiac myocyte function. The proposed experiments, using state-of-the-art electrophysiology and optical methods, as well as biochemical assays, will evaluate the effects of NOS1 and NOS3 on cardiac myocyte function. The results of these studies will provide a new integrated understanding of how NOS1 and NOS3 regulate cardiac myocyte function and will help delineate their role in cardiac disease states. The specific aims are designed to examine the mechanisms, end targets and signaling pathways of NOS1 and NOS3. The hypothesis is that NOS1 and NOS3 differentially regulate cardiac contractility by ultimately modulating sarcoplasmic reticulum Ca2+ load through distinct end targets and signaling pathways. The specific aims are to: 1) define physiological effects of NOS1 and NOS3 on cardiac myocyte function; 2) determine mechanisms of NOS1 and NOS3 by examining specific end targets; 3) elucidate the downstream pathways of NOS1 or NOS3 on sarcoplasmic reticulum Ca2+ ATPase, phospholamban and the L-type Ca2+ channel. Studies will be performed on myocytes and trabeculae isolated from wildtype, NOS1 knockout, NOS3 knockout and phospholamban knockout mice, as well as rabbits. Heart disease is one of the leading causes of mortality in the Western world. The relevance of this proposal to public health is that these results may improve the treatment of heart disease by potentially identifying new targets for drug development.

描述（由申请人提供）：心肌细胞组成型表达称为神经元氧化物合酶（NNOS，NOS1）和内皮一氧化氮合酶（ENOS，NOS3）的酶。通过NOS1和NOS3产生的一氧化氮（NO）是正常健康心脏中心脏收缩性的重要调节剂。 NOS1和NOS3在患病心脏的重塑和心脏功能障碍中也起着关键作用。已经发现NOS1对于阳性力频率反应和增强β-肾上腺素能反应至关重要。这是有争议的，因为其他研究发现结果有些不同。可以更好地理解NOS3的效果，因为NOS3限制了β-肾上腺素能反应。但是，尚不清楚NOS1和NOS3的确定机制，最终目标和信号通路。因此，该提案的目的是批判性地检查NOS1和NOS3如何调节心肌细胞功能。提出的使用最先进的电生理学和光学方法以及生化测定的实验将评估NOS1和NOS3对心肌细胞功能的影响。这些研究的结果将对NOS1和NOS3如何调节心肌细胞功能有了新的综合了解，并将有助于描述其在心脏病状态中的作用。该特定目的旨在检查NOS1和NOS3的机制，最终目标和信号通路。假设是NOS1和NOS3通过最终通过不同的最终目标和信号通路调节肌质网Ca2+负载来差异调节心脏收缩。具体目的是：1）定义NOS1和NOS3对心肌细胞功能的生理影响； 2）通过检查特定最终目标来确定NOS1和NOS3的机制； 3）阐明肌质网Ca2+ ATPase，Phospholamban和L型Ca2+通道上NOS1或NOS3的下游途径。研究将对从野生型，NOS1敲除，NOS3敲除和磷脂敲除小鼠以及兔子分离的心肌细胞和小梁进行研究。心脏病是西方世界死亡率的主要原因之一。该提议与公共卫生的相关性是，这些结果可以通过识别新的药物开发目标来改善心脏病的治疗。