Cardiac Physiology and Imaging Core

摘要 心脏生理学和成像核心（核心C）将为所有4个项目提供服务，目标是 产生人胚胎干细胞衍生的心肌细胞（hESC-1）的功能和结构分析。 CM;目标1）和成年小鼠心脏（目标2）。具体而言，目标1中使用的测定将提供高质量 hESC-CM结构和功能的无偏评估。Core C的服务提高了 项目通过提供一致的专业知识，在功能和结构评价的hESC-CM使用 多种互补技术。其中包括用于结构评估的共聚焦荧光显微镜 以及细胞溶质[Ca 2 +]和膜电压成像，一种用于多细胞 电生理和收缩评估，透射电子显微镜（TEM）用于超微结构 分析，用于单个hESC-CM电生理测量的膜片钳技术，和原位 杂交（RNAscope）用于评估空间基因表达模式。项目1受益于核心C， 单细胞水平的基因表达分析。项目2和项目3在数量上受益于核心C hESC-CM的电和收缩性质的评估。所有项目都受益于量化 hESC-CM的肌节组织、极性和形状的分析。目标2中使用的技术将 为成年小鼠心脏提供高质量无偏倚的功能和结构评估。核心C服务 包括小鼠超高频超声、麻醉小鼠体表ECG、遥测ECG监测 的清醒小鼠，离体小鼠心脏的光学电压/钙标测，以及 心肌细胞S期活动使用定制的成像系统。核心将生成高分辨率 项目3中的超声图像，以提供与以下相关的结构、功能和血流的定量数据： 心脏发育中的基因改变ECG和电压/钙标测将提供以下定量 在项目2和3中的心脏电功能。心肌细胞S期活动的定量将有助于 评估所有项目中拟议的基因改变对心肌细胞增殖的影响。

Abstract The Cardiac Physiology and Imaging Core (Core C) will provide service to all 4 projects with the goals of generating functional and structural analyses of human Embryonic Stem Cell-derived cardiomyocytes (hESC- CMs; Aim 1) and the adult mouse heart (Aim 2). Specifically, assays utilized in Aim 1 will provide high quality unbiased assessment of hESC-CM structure and function. Core C’s services enhance the efficiency of the Projects by providing consistent expertise in the functional and structural evaluation of hESC-CMs using a variety of complementary techniques. These include confocal fluorescence microscopy for structural evaluation as well as cytosolic [Ca2+] and membrane voltage imaging, an electrode array system for multicellular electrophysiological and contractile assessment, Transmission Electron Microscopy (TEM) for ultrastructural analyses, the patch-clamp technique for single hESC-CM electrophysiological measurements, and in situ hybridization (RNAscope) for evaluation of spatial gene expression patterns. Project 1 benefits from Core C in the analyses of gene expression at the single cell level. Projects 2 and 3 benefit from Core C in the quantitative assessment of electrical and contractile properties of hESC-CMs. All Projects benefit from the quantitative analysis of sarcomeric organization, polarity and shape of hESC-CMs. Techniques to be used in Aim 2 will provide high quality unbiased functional and structural assessment of the adult mouse heart. Core C’s services include ultra-high frequency ultrasound of mice, surface ECG in anesthetized mice, telemetric ECG monitoring of conscious mice, optical voltage / calcium mapping of isolated mouse hearts, and quantitation of cardiomyocyte S-phase activity using a custom-built imaging system. The Core will generate high-resolution ultrasound images in Projects 3 in order to provide quantitative data of structure, function, and flow relative to genetic alterations in cardiac development. ECG and voltage / calcium mapping will provide quantitation of cardiac electrical function in Projects 2 and 3. Quantitation of cardiomyocyte S-phase activity will aid in assessing effects of proposed genetic alterations on cardiomyocyte proliferation in all Projects.