Decreased Cholinergic Tone and Mitochondrial Dysfunction in Heart

心脏胆碱能张力降低和线粒体功能障碍

• 批准号: 8327739
• 负责人: William Jonathan Lederer
• 金额: $ 5.27万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-02 至 2014-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8327739
• 关键词: Affect; Aging; Baltimore; Behavior; Biology; Biomedical Engineering; Biomedical Technology; Brazil; Calcium; Cardiac; Cardiac Myocytes; Cell physiology; Cells; Cellular biology; Cholinesterase Inhibitors; Chronic; Collaborations; Complement; Complex; Data; Deletion Mutation; Disease; Dose; Employee Strikes; Enzymes; Equilibrium; Functional disorder; Funding; Gene Expression; Grant; Head; Heart; Heart Diseases; Heart failure; Image; Instruction; Intervention; Ion Channel; Kinetics; Learning; Macromolecular Complexes; Maryland; Measures; Membrane; Membrane Potentials; Methods; Mitochondria; Molecular; Molecular and Cellular Biology; Mus; Muscle Cells; Mutant Strains Mice; Myocardial; Nervous system structure; Physiology; Play; Process; Production; Reactive Oxygen Species; Regulation; Research; Research Personnel; Resolution; Role; RyR2; Sarcolemma; Signal Transduction; Site; Techniques; Testing; Time; Training; United States National Institutes of Health; Universities; Ventricular; Work; acetylcholine transporter; base; cell behavior; cholinergic; heart cell; heart function; heart rhythm; human morbidity; human mortality; improved; mitochondrial dysfunction; mutant; new therapeutic target; novel; parent grant; patch clamp; pyridostigmine; research study; restoration; sudden cardiac death

DESCRIPTION (provided by applicant): This research will be done primarily in Brazil at Universidade Federal de Minas Gerais in collaboration with Dr. Silvia Guatimosim, as an extension of Project 3 of NIH Grant number P01 HL67849 (A.R. Marks), 4/1/2006-3/31/2011. Dr. W. Jonathan Lederer is the project leader. The proposed FIRCA project seeks to enable novel real-time imaging and cell biology experiments for an outstanding research group in Brazil headed by Dr. Silvia Guatimosim. The new work will be made possible by using the PI's state-of-the-art facilities at the University of Maryland Center for Biomedical Engineering and Technology to train Dr. Guatimosim and her colleagues in new methods. Broadly the co-investigators will investigate how the cholinergic tone in heart affects Ca2+ signaling in the cardiac myocytes and mitochondrial function. Imaging and biophysical methods will be used along with a novel mouse line (VaChT KDHOM mice) that has reduced expression of the vesicular acetylcholine transporter. This novel mouse line was developed with previous FIRCA funding to Dr. M. Prado and will permit the co-investigators to characterize the consequences of cholinergic hypofunction on heart cell behavior. Provocative preliminary results by Dr. Guatimosim show that the VAChT KDHOM mouse has heart failure including decreased myocardial force, altered ventricular calcium handling and molecular remodeling (Lara et al., Molecular & Cellular Biology, 2010 in press), including altered mitochondrial biology and enzyme levels and increased production of reactive oxygen species (ROS) (see Preliminary Results). Much of this dysfunction was reversed by treatment with a cholinesterase inhibitor (pyridostigmine). Since mitochondria are considered the powerhouse of the cell, the central hypothesis of this FIRCA project is that alterations in mitochondrial dynamics contribute to cardiac malfunction in VAChT mutant mice. To test this hypothesis, we will measure mitochondrial dynamics in patch-clamped ventricular myocytes from VAChT mutants by using a combination of real-time imaging of mitochondrial Ca2+ levels and membrane potential ([Ca2+]mito )The functional data obtained by real-time imaging will provide an integrated understanding of mitochondria' s role on heart disease caused by reduced cholinergic tone. This FIRCA proposal will provide the means to build new research capabilities at the Universidade Federal de Minas Gerais site for the simultaneous patch-clamp and real-time imaging of mitochondria, and to incorporate these state-of-the-art techniques into Dr. Guatimosim's research. Dr. Lederer will provide training in Baltimore for Dr. Guatimosim and her co-workers and on-site instruction in Brazil.

描述（由申请人提供）：这项研究将主要在巴西与米纳斯·格拉斯大学（Universidade Federal de Minas Gerais）与Silvia Guatimosim博士合作，作为NIH赠款编号P01 HL67849（A.R. Marks）的项目3的扩展，4/1/1/2006-3/31/2011。 W. Jonathan Lederer博士是项目负责人。拟议的FIRCA项目旨在为巴西杰出的研究小组Silvia Guatimosim博士领导的新型实时成像和细胞生物学实验。通过在马里兰州生物医学工程技术中心使用PI的最先进设施来培训Guatimosim博士及其同事以新方法来培训新作品。从广义上讲，共染色器将研究心脏中的胆碱能张力如何影响心肌细胞和线粒体功能中的Ca2+信号传导。成像和生物物理方法将与一种新型小鼠系（Vacht KDHOM小鼠）一起使用，该系列降低了囊泡乙酰胆碱转运蛋白的表达。这条新颖的小鼠系列是先前向M. Prado博士提供的FIRCA资助，并将允许共同研究者表征胆碱能功能低下对心脏细胞行为的后果。 Provocative preliminary results by Dr. Guatimosim show that the VAChT KDHOM mouse has heart failure including decreased myocardial force, altered ventricular calcium handling and molecular remodeling (Lara et al., Molecular & Cellular Biology, 2010 in press), including altered mitochondrial biology and enzyme levels and increased production of reactive oxygen species (ROS) (see Preliminary Results).这种功能障碍的大部分是通过胆碱酯酶抑制剂（Pyridostigmine）治疗逆转的。由于线粒体被认为是细胞的动力体，因此该FIRCA项目的中心假设是线粒体动力学的改变会导致VACHT突变小鼠的心脏故障。为了检验该假设，我们将通过使用对线粒体Ca2+水平的实时成像的组合（[CA2+] mito）组合通过对实时成像获得的功能数据，可以通过对心脏方位的一部分，通过实时成像的理解，对线粒体成像的理解会加强心脏方位的方法。这项FIRCA提案将提供在联邦De Minas Gerais网站上建立新的研究能力的方法，以同时进行线粒体和实时成像，并将这些最先进的技术纳入Guatimosim博士的研究中。 Lederer博士将在巴尔的摩为瓜蒂莫森博士及其同事以及巴西的现场指导提供培训。