LEICA TCS SP CONFOCAL MICROSCOPE

该提案请求支持购买Al Leica TCS SP共焦显微镜。这台显微镜将成为共享资源，对于在纽约大学医学中心几个部门工作的研究人员正在进行的研究至关重要。主要用户的项目侧重于分析细胞间的相互作用和细胞内的信号通路，并要求在培养的细胞或复杂组织中对蛋白质或细胞成分进行精确定位。大多数项目都集中在发育中的神经系统中细胞与细胞的相互作用上。这些研究包括：i)转基因小鼠神经肌肉突触形成的调控和调控突触后蛋白聚集的机制(Burden)，ii)通过共定位培养细胞中的信号成分来调控胶质形成的信号通路(Chao，Salzer)和分析转基因小鼠(Chao)神经胶质发育的研究，iii)细胞黏附分子和其他信号，调控线虫(Clark)中表达GFP的特定神经元和果蝇(Sink)出口交界处运动纤维向后定向突起的生长，以及iv)哺乳动物中枢神经系统中发育中神经元的分化调控机制。第二个研究领域涉及宿主细胞/寄生虫的相互作用，研究志贺氏菌和沙门氏菌通过激活ICE依赖途径(Zychlinsky)导致巨噬细胞凋亡的机制。将蛋白质精确定位到特定的细胞隔间并通过共定位推断蛋白质相互作用的能力对于理解致病机制和信号转导机制至关重要。上述项目是由资深和新近招聘的教职员工承担的，他们将合作支持和运营这一共焦设施。这种显微镜的分辨率和灵敏度的提高，以及它对较厚组织有效成像的能力，将是这些项目成功的关键。

This proposal request support for the purchase of al Leica TCS SP confocal microscope. This microscope, which will be a shared resource, will be crucial for the ongoing research of investigators working in several departments at NYUMC. The projects of the major users are focused on analyzing cell-cell interactions and intracellular signaling pathways and require precise localization of proteins or cellular constituents in cultured cells or complex tissues. Most projects are focused on cell-cell interactions in the developing nervous system. These include studies of: i) the regulation of neuromuscular synapse formation in genetically modified mice and the mechanisms that regulate clustering of postsynaptic proteins (Burden), ii) the signaling pathways that regulate gliogenesis by co- localizing signaling components in cultured cells (Chao, Salzer) and analyzing glial development in genetically modified mice (Chao), iii) the cell adhesion molecules and other signals that regulate the posteriorly- directed outgrowth of specific GFP expressing neurons in C. elegans (Clark) and motor fibers at the exit junction in Drosophila (Sink), and iv) the mechanisms that regulate differentiation of developing neurons in the mammalian central nervous system (Fishell). A second area of research, which is concerned with host cell/parasite interactions, studies the mechanisms by which Shigella and Salmonella lead t macrophage apoptosis via activation of ICE-dependent pathways (Zychlinsky). The ability to precisely localize proteins to specific cellular compartments and to infer protein intera9tions by co-localization will be crucial to an understanding of the pathogenetic and signal transduction mechanisms. The above projects are being undertaken by senior and recently recruited faculty members, who will cooperate in supporting and operating this confocal facility. The enhanced resolution and sensitivity of this microscope, as well as its ability to effectively image thicker tissue, will be crucial to the success of these projects.