BIOMEDICAL RESEARCH SUPPORT GRANT

现代细胞生物学最重要的努力之一是 了解细胞间和细胞内的信号传递机制。 目前我们的实验室正在深入研究的著名例子 包括：1)离子通道和神经递质的分化 胚胎神经元中的受体；2)免疫细胞与 抗原及其靶细胞；3)突触发育 神经系统中的接触；4)细胞-细胞连接的调节 耦合；5)胞内包装和定向转运 6)神经过程中细胞表型的分配 发展。一群代表以下领域的教职员工 Interest将结合他们的专业知识开发一个基于 激光扫描共聚焦显微镜和视频成像。最近的一部分 细胞生物学的革命是由于荧光技术的发展 使用计算机增强显微镜可视化的指示剂染料 技巧。这些染料允许对细胞内进行定量测量 离子、信使分子、膜电位和特异性标记 活细胞的膜和细胞内成分与时间和 能够忠实地监测地点和时间的空间分辨率 细胞内事件的进程。我们计划发展下一代 基于显微镜的视频采集和分析系统，我们使用它 将研究第二信使的三维分布 细胞内分子的原位，以及快速变化的动力学 转导事件。这个新系统将结合改进的光学系统， 三维成像能力，以及能够 实时获取、分析和存储处理后的图像。我们会 结合膜片钳技术，实现构象的解析 单离子通道蛋白在测量时的变化 细胞内信使分子的三维分布。 笼状化合物的光解将允许快速改变 活跃的第二信使集中。这个设施将极大地 增强现有能力，进一步开展细胞- 信令机制。

One of the most significant endeavors of modern cell biology is to understand mechanisms that mediate signalling between and within cells. Notable examples presently under intensive study in our laboratories include: 1) the differentiation of ion channels and neurotransmitter receptors in embryonic neurons; 2) the interactions of immune cells with antigens and with their target cells; 3) the development of synaptic contacts in neural systems; 4) the regulation of cell-cell junctional coupling; 5) the packaging and directed transport of intracellular components; and 6) the assignment of cell phenotype during neural development. A group of faculty members representing these areas of interest will combine their expertise to develop a facility based on a laser-scanning confocal microscope and video imaging. Part of the recent revolution in cell biology is due to the development of fluorescent indicator dyes that are visualized using computer-enhanced microscopy techniques. These dyes permit quantitative measurement of intracellular ions, messenger molecules, membrane potential, and specific labeled membrane and intracellular constituents of living cells with temporal and spatial resolutions capable of faithfully monitoring the sites and time courses of intracellular events. We propose to develop the next generation of microscope-based video acquisition and analysis system with which we will investigate the three-dimensional distributions of second messenger molecules in cells in situ, and the kinetics of rapidly changing transduction events. This new system will incorporate improved optics, three-dimensional imaging capabilities, and a detector system capable of acquiring, analyzing, and storing processed images in real time. We will incorporate patch-clamp techniques, enabling resolution of conformational changes in single ion-channel proteins while measuring the three-dimensional distributions of intracellular messenger molecules. Photolysis of caged compounds will allow for rapid changes in the concentration of active second messengers. This facility will greatly enhance the existing capabilities and further the investigation of cell- signalling mechanisms.