Cellular Fluorescence-Contractility Imaging System

描述(由申请人提供)：新英格兰医学中心的分子心脏病研究所(MCRI)致力于探索人类心血管疾病的分子机制。MCRI研究人员使用体外分子生物学、细胞生物学和转基因方法来识别和验证新的治疗靶点。MCRI内的研究人员寻求不同但互补的研究途径--其中绝大多数是由NIH支持的。MCRI的两大研究领域包括血管生物学和心肌细胞生物学。我们研究血管紧张性、心肌细胞肥大和细胞凋亡、心脏发育和细胞增殖的正常和病理生理调节和功能。对于许多由NIH资助的MCRI研究人员--包括这项提案中的调查人员--来说，一个共同的焦点是调节心脏和血管肌细胞收缩和松弛的机制。为了了解调控心肌细胞和血管平滑肌细胞收缩和松弛的分子事件，我们对培养的心脏和血管细胞进行了广泛的细胞和生化事件的研究，这些细胞和生化事件是在我们的细胞培养设施中常规地产生、传代和储存的。这是一个研究心血管信号的有效系统，最大限度地减少了对完整动物的使用。然而，目前我们还没有能力对单个细胞中的细胞信号和细胞收缩和松弛提供复杂的生理学分析。因此，MCRI确定了对成像设备的需求，该设备可以同时测量细胞收缩和松弛，并实时执行高分辨率荧光成像。这些目标集中在两个领域获得新技术：与细胞收缩相关的钙成像，以及信号转导途径和蛋白质的荧光检测，以研究其活性和细胞内定位。 因此，基于MCRI正在进行的计划和对心脏和血管细胞生理学研究能力的日益增长的需求，这项建议的目的是获得所需的支持，以建立一个高度集成的细胞成像系统，以测量实时的单细胞收缩松弛和同时的钙瞬变。收购这一系统将促进我们机构内对心血管信号转导的广泛和补充研究。

DESCRIPTION (provided by applicant): The Molecular Cardiology Research Institute (MCRI) at the New England Medical Center is devoted to the exploration of the molecular mechanisms that underlie human cardiovascular diseases. MCRI investigators use in vitro molecular biological, cell biological and transgenic approaches to identify and validate new therapeutic targets. The investigators within the MCRI pursue varied, yet complementary avenues of research--the vast majority of which are NIH-supported. Two broad areas of research within the MCRI include vascular and cardiomyocyte biology. We examine the normal and pathophysiologic regulation and function of vasomotor tone, cardiomyocyte hypertrophy and apoptosis, cardiac development and cell proliferation. A common focus for many of the NIH-funded MCRI Investigators--those who comprise the Investigators in this proposal--is mechanisms that regulate contraction and relaxation of cardiac and vascular myocytes. In order to understand the molecular events that regulate cardiac myocyte and vascular smooth muscle cell contraction and relaxation, cellular and biochemical events are studied extensively in cultured cardiac and vascular cells, which are routinely generated, passaged and stored in our cell culture facilities. This is an efficient system for the study of cardiovascular signaling that minimizes the use of intact animals. However, at present we do not have the capacity to provide sophisticated physiological analysis of cell signaling and cell contraction and relaxation in single cells. The MCRI has thus identified the need for an imaging apparatus that can simultaneously measure cell contraction and relaxation and perform highresolution fluorescence imaging in real-time. These goals are focused on acquiring new technology in two areas: calcium imaging related to cell contraction, and signal transduction pathways and fluorescence detection of proteins to study their activity and intracellular localization. Thus, based on ongoing programs and increasing needs within the MCRI for the capacity to study cardiac and vascular cell physiology, the purpose of this proposal is to gain the support needed to build a highly integrated cell imaging system to measure real-time single cell contraction relaxation and simultaneous calcium transients. Acquisition of this system will facilitate the extensive and complementary studies of cardiovascular signal transduction within our institution.