Development of a set up for the combined measurement of force, myocyte tension and mitochondrial redox status in rat engineered heart tissue to characterize cardiac hypertro

该项目的目的是建立一个新的设置，允许在电刺激工程心脏组织中测量心脏生物学中的两个重要参数-肌细胞张力（作为整个心脏壁应力的替代品）和线粒体氧化还原状态。两者都将采用基于荧光的技术进行测量，同时测量人工心肌的发育力。对于肌细胞紧张，将建立一个基于fret的成像系统，可视化机械敏感的apelin受体和β -arrestine之间的相互作用。一种带有线粒体识别序列的修饰红色荧光蛋白将用于可视化线粒体氧化还原状态。该系统将测量不同负荷前后的影响，以及肌丝钙敏感性的药理修饰对心肌细胞张力和线粒体氧化还原状态的影响。壁应激和线粒体氧化还原状态可能在心脏肥大和遗传决定的心肌病的发病机制中发挥重要作用，但仍未完全理解。该项目的成功结果可能会对导致心脏肥厚的这些重要生物过程产生新的见解。

The aim of this project is the establishment of a new setup that allows the measurement of two important parameters in heart biology in electrically stimulated engineered heart tissues - myocyte tension (as a surrogate for wall stress in whole hearts) and mitochondrial redox status. Both will be measured on a fluorescence-based technique in parallel to the developed force of the artificial heart muscle. For myocyte tension, a FRET-based imaging system that visualizes the interaction between the mechanosensitive apelin receptor and beta-arrestine will be established. A modified red fluorescent protein with a mitochondrial recognition sequence will be used to visualize the mitochondrial redox status. The system will provide measurements of the influence of different pre- and afterload as well as pharmacological modification of myofilament calcium sensitivity on myocyte tension and the mitochondrial redox status in work-performing heart muscles. Wall stress and mitochondrial redox status likely play an important, but still incompletely understood role in the pathogenesis of cardiac hypertrophy and genetically determined cardiomyopathies. The successful outcome of this project might lead to novel insights in these important biological processes leading to cardiac hypertrophy.