Myocyte Isolation and Myocyte and Cardiac Physiology

项目总结(见说明)： 心脏和心肌细胞生理学核心D的目的是为评估糖共轭修饰对心肌细胞、肌肉纤维和完整心脏生理学的影响提供必要的方法。采用两种体外方法和两种非侵入性体内方法。对于项目1、2和3，我们将使用分离的成人心肌细胞来确定蛋白质修饰对肌节缩短和松弛的影响，以及对整个细胞钙瞬变的影响。对静息和肾上腺素能刺激细胞进行了研究。成人心脏模型的心肌细胞分离也被提供给该项目，用于分子研究和线粒体的分离。对于项目2和项目3，我们将提供离休心脏朗宁多夫制剂，以确定器官功能，并提供组织用于使用缺血/再灌注和预适应模型进行额外分析。对于项目1和项目5，核心将使用FIME线路上的非侵入性回波-多普勒成像分析来确定所使用的动物模型的心脏功能。不同饮食的小鼠动脉粥样硬化模型是这项分析的主要焦点。最后，对于项目2和项目3，我们将使用两个位置的主动脉血流的多普勒成像和脉搏波速度的测定来确定血管特性，例如大动脉僵硬。我们有各种各样的最先进的方法来评估心脏功能，一些具体的研究将取决于获得的最终数据。

PROJECT SUMMARY (See instructions): The purpose of the Cardiac and Myocyte Physiology Core D is to provide required methods for the evaluation of the effects of glycoconjugate modifications on myocyte, muscle fiber, and intact heart physiology. Two in vitro methods and two non-invasive in vivo methods are employed. For Projects 1, 2 and 3, we will use isolated adult myocytes to determine the impact of protein modification on sarcomere shortening and relaxation, and on the whole cell calcium transient. Resting and adrenergic stimulated cells are studied. Myocyte isolation from adult heart models also are provided to the project for molecular studies and for the isolation of mitochondria. For Projects 2 and 3 we will provide isolated heart Langendorff preparations, to define organ function, and to provide tissue for additional analysis using models of ischemia/reperfusion and preconditioning. For Projects 1 and 5, the Core will determine cardiac function using non-invasive echo-Doppler imaging analysis over a fime-line for the animal models used. Mouse models of atherosclerosis on varying diets are a major focus of this analysis. Lastly, for Projects 2 and 3, we will determine vascular properties such as large artery stiffness using Doppler imaging of aortic flow at two locations and determination of pulse wave velocity. We have a wide variety of state-of-the-art methods to evaluate cardiac functions and some of the specific studies will be determined by the inifial data obtained.