Regulatory Mechanisms in Lymphatic Muscle Contraction

淋巴肌收缩的调节机制

• 批准号: 8105099
• 负责人: MARIAPPAN MUTHUCHAMY
• 金额: $ 10.04万
• 依托单位: TEXAS A&M UNIVERSITY HEALTH SCIENCE CTR
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8105099
• 关键词: Actins; Address; Adrenergic Agonists; Affect; Animals; Bathing; Behavior; Binding; Blood Circulation; Body Fluids; Body Regions; Bradykinin; Buffers; Calcium; Caliber; Cardiac; Cardiovascular system; Cells; Central Vein; Characteristics; Chest; Clinical; Complement; Contractile Proteins; Data; Depressed mood; Disease; Down-Regulation; Edema; Equilibrium; Exhibits; Functional disorder; Gene Expression; Gene Knock-Out Model; Generations; Genes; Genomics; Goals; Immune; Immunoblotting; Immunofluorescence Immunologic; Immunoprecipitation; Inflammation Mediators; Injury; Isometric Contraction; Japan; Knock-out; Knockout Mice; Laboratories; Levosimendan; Lymph; Lymphatic; Lymphatic System; Lymphatic vessel; Lymphocyte; Measurement; Measures; Mediating; Mesentery; Methods; Microfilaments; Molecular; Molecular Genetics; Molecular Profiling; Monitor; Motor; Mus; Muscle; Muscle Contraction; Muscle Proteins; Muscle function; Myosin Heavy Chains; Myosin Light Chain Kinase; Myosin Light Chains; Nature; Peptides; Pharmaceutical Preparations; Phosphorylation; Physiological; Play; Preparation; Protein Isoforms; Proteins; Proteomics; RNA; Rattus; Regulation; Regulatory Element; Regulatory Pathway; Research; Research Project Grants; Role; Saline; Series; Signal Pathway; Skin; Small Interfering RNA; Smooth Muscle; Smooth Muscle Myosins; Striated Muscles; Students; Substance P; System; Temperature; Testing; Thick Filament; Thin Filament; Thoracic Duct; Tissues; Training; Transducers; Transfection; Transgenic Mice; Transgenic Model; Tropomyosin; Troponin; Troponin C; Tungsten; United States National Institutes of Health; Vascular Smooth Muscle; Visit; Western Blotting; abstracting; base; caldesmon; calponin; career; genetic regulatory protein; immune function; kinase inhibitor; knock-down; knowledge base; lipid metabolism; lymph flow; mRNA Expression; mouse model; neoplastic cell; non-muscle myosin heavy chain-B; overexpression; pressure; promoter; research study; response; synthetic peptide; therapy development; tool

DESCRIPTION (provided by applicant): My immediate career objectives are to develop a reputation for high quality research and to train graduate and postdoctoral students in the cardiovascular and lymphatic research field. My long-term career are to advance the level of understanding the regulatory mechanisms of lymphatic muscle contractility under normal and diseased/pathophysiological conditions. Lymphatic muscle exhibits strong/phasic contractions, much higher shortening velocities and different intracellular calcium dynamics. Unfortunately little information is known about the molecular mechanisms that are responsible for these unique characteristics. In this proposal we will use the available contractile protein gene knock out models or transgenic models with overexpression of contractile protein isoforms to test the central hypothesis remodeling of thick or thin filament proteins in the contractile apparatus of lymphatic muscle modulates its contractile dynamics through altering the calcium sensitivity and crossbridge activation mechanisms. The specific aims are: (1) To determine the functional roles of SM-B myosin heavy chain (MHC) in the phasic and tonic contraction of lymphatics, (2) To define the roles of tropomyosin in the thin filament-mediated contraction of lymphatics. We will use SM-B MHC knockout and SM-MHC/calponin double knockout mouse models. The vascular smooth muscle a-actin promoter will be to express striated muscle tropomyosin in lymphatic muscle. An adenoviral siRNA approach will be used knock down the smooth muscle a- and p-TM gene expression in lymphatics. We will isolated/cannulated vessels from iliac and thoracic duct lymphatics to study the contractile characteristics of lymphatics. Force and calcium measurements will be conducted in the lymphatic segment preparations (both intact and skinned) to determine the calcium sensitivity and cooperativity mechanisms of lymphatics. The contractile mechanisms of lymphatics are poorly understood and this study will significantly advance our knowledge of the basis for the lymphatic vessel function. These studies further advance my training lymphatic research using mouse models, which would allow me to develop and use genomic proteomic approaches to determine the signaling pathways that regulate lymphatic muscle function. (End of Abstract)

描述（由申请人提供）： 我的直接职业目标是在心血管和淋巴研究领域培训高质量研究的声誉，并培训毕业生和博士后学生。我的长期职业是提高理解在正常和患病/病理生理状况下淋巴肌肉收缩力的调节机制的水平。淋巴肌肉表现出强/阶段的收缩，较高的缩短速度和不同的细胞内钙动力学。不幸的是，关于这些独特特征负责的分子机制知之甚少。在此提案中，我们将使用具有收缩蛋白同工型过表达的可用收缩蛋白基因敲除模型或转基因模型来测试淋巴肌肉收缩型厚或薄丝蛋白的中心假设重塑，从而通过改变钙化肌肉动力学来改变钙化动力学。具体目的是：（1）确定SM-B肌球蛋白重链（MHC）在淋巴管的阶段和滋补性收缩中的功能作用，（2）定义了对tropomyosin在淋巴管的薄丝介导的淋巴管的缩写中的作用。我们将使用SM-B MHC敲除和SM-MHC/Calponin双基因敲除鼠标模型。血管平滑肌A-肌动蛋白启动子将在淋巴肌中表达横纹肌肉肌球蛋白。将使用一种腺病毒siRNA方法击倒淋巴管中平滑肌A-和P-TM基因表达。我们将从iLiac和胸管淋巴管中分离/插管的血管，以研究淋巴管的收缩特征。力和钙测量将在淋巴段制剂（完整和皮肤）中进行，以确定淋巴管的钙敏感性和协同机制。淋巴管的收缩机制知之甚少，这项研究将显着提高我们对淋巴管功能基础的了解。这些研究进一步推进了我使用小鼠模型的训练淋巴研究，这将使我能够开发和使用基因组蛋白质组学方法来确定调节淋巴肌肉功能的信号传导途径。 （抽象的结尾）