Luminescence Imaging: Molecules, Cells and Tissues

发光成像：分子、细胞和组织

• 批准号: 7047193
• 负责人: David C Spray
• 金额: $ 20.88万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-05-15 至 2009-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7047193
• 关键词: astrocytes; astrocytoma; bone marrow; cytokine; electromagnetic radiation; gap junctions; gene expression; genes; housing; light intensity; luminescence; medicine; membrane channels; tissue /cell culture; tissues; training; transplantation; university

DESCRIPTION (provided by applicant): The purpose of this application is to obtain a photon-counting camera and accessories to allow high temporal and spatial resolution of luminescence within cultured cells and in whole animals; accessories include both multiwell plate and single tube luminometers required for optimization of expression and activation of the photoproteins in vitro and an imaging station in which whole animal imaging will be performed. There is no such high sensitivity camera or whole animal imager currently at Einstein College of Medicine, and there is currently neither type of luminometer in the nine-floor Kennedy Center housing the Department of Neuroscience, where the proposed studies will be performed. The studies that are proposed are all natural extensions of NIH-funded projects of nine investigators that will use the relatively new technique of spatially resolved luminescence recording that has become more accessible due to new generations of vector constructs in which the photoproteins have been optimized with regard to intensity and wavelengths of light output and destabilized with regard to gene expression reporting. The studies of users outlined in this proposal are diverse, including bioluminescent resonance energy transfer to determine dimerization and binding partners for connexin molecules, luminescent reporter molecules to localize ATP release from within astrocytes and astrocytoma cells, luminescence-tagged bone marrow cells in order to characterize their tissue distribution following transplantation into mice, and gene promoters fused to luminescent reporter molecules to identify time course of gene activation in cytokine-stimulated astrocytes and to develop a moderately high throughput assay by which to identify connexin-specific gap junction channel blockers. The proposal also includes the plan for a bimonthly user group meeting, through which new users will be attracted to the instrument and new analytical tools and protocols will be discussed, as well as problems that may arise. It also includes plans for training as well as inclusion of relevant methods in a graduate student course.

描述（由申请人提供）：本申请的目的是获得光子计数照相机和附件，以实现培养细胞和整个动物中发光的高时间和空间分辨率;附件包括优化体外发光蛋白表达和激活所需的多孔板和单管光度计以及将进行整个动物成像的成像站。目前爱因斯坦医学院还没有这样的高灵敏度相机或整体动物成像仪，目前在神经科学系九层楼的肯尼迪中心也没有这两种类型的光度计，拟议的研究将在那里进行。提出的研究都是NIH资助的9名研究人员的项目的自然延伸，这些研究将使用相对较新的空间分辨发光记录技术，由于新一代载体构建，该技术已变得更容易获得，其中光蛋白在光输出的强度和波长方面进行了优化，并且在基因表达报告方面不稳定。本提案中概述的用户研究多种多样，包括生物发光共振能量转移以确定连接蛋白分子的二聚化和结合伴侣，发光报告分子以定位星形胶质细胞和星形细胞瘤细胞内的ATP释放，发光标记的骨髓细胞以表征其移植到小鼠后的组织分布，和与发光报道分子融合的基因启动子，以鉴定精氨酸刺激的星形胶质细胞中基因激活的时间过程，并开发一种中等高通量的测定，通过该测定鉴定连接蛋白特异性间隙连接通道阻断剂。该提案还包括每两个月举行一次用户小组会议的计划，通过该会议将吸引新用户使用该仪器，并将讨论新的分析工具和协议以及可能出现的问题。它还包括培训计划以及将有关方法纳入研究生课程。

项目成果

Pannexin 1 channels play essential roles in urothelial mechanotransduction and intercellular signaling.

• DOI: 10.1371/journal.pone.0106269
• 发表时间: 2014
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Negoro H;Urban-Maldonado M;Liou LS;Spray DC;Thi MM;Suadicani SO
• 通讯作者: Suadicani SO