CORE--NEUROCYTOLOGY /CELLULAR IMAGING

宗旨和目标 遗传性和环境性发育障碍可能会对 中枢神经系统(CMS)的发育和功能。其发病机制和致病机理 神经修复以及翻译研究将导致 损伤的中枢神经系统再生的治疗方法日益成为研究的主题 包括本中心成员在内的研究人员的紧张研究。与复杂性相匹配 在这些问题中，一个人需要多学科的方法。因此，它将对我们的 神经科学家来到一个单一的综合神经科学核心，在那里他们可以得到建议 由专家教职员工组成的团队使用适当的方法、技术和设备，以 调查他们的神经科学研究问题。为了实现这一目标，有3个核心 合并(细胞生物学、细胞学和成像)。昂贵的最先进的仪器，如 蔡司激光扫描共聚焦显微镜LSM 510 META和徕卡激光细胞捕获 购买显微解剖系统。额外的教员被招募来提供广泛的 神经科学专业知识，以及其他MRRC和校园提供的贡献 这将令人满意地满足核心用户表所列项目的巨大需求。 这里将介绍所提供服务的基本原理。 神经科学和成像核心提供专业知识、服务和设备 支持细胞和分子神经科学研究： 干细胞/细胞培养/细胞移植 *斑马鱼设施 ？神经细胞学 ？免疫细胞化学 原位杂交 ？共焦显微镜(蔡司510META) ？从组织切片和细胞培养中捕获激光细胞 首先，细胞培养和细胞移植的体外/体内方法将是 讨论过。发现成年中枢神经系统具有通过多潜能再生的潜力 干细胞，引起了人们对研究神经干细胞的日益浓厚的兴趣 在动物模型中理解神经退行性疾病的发病机制 退行性疾病的治疗工具。此核心的目的是满足以下需求 融入新技术，旨在研究中枢神经系统功能的恢复。《核心》 结合了不同的专业领域，包括特定神经细胞的培养，神经干细胞 和ES细胞的增殖、适当分化、鉴定及其在神经中的应用 移植。这些方法已被证明在中枢神经系统修复领域很有前途。这个 CORE整合了最先进的胚胎干细胞制备方法 神经干细胞，以及神经元和神经胶质细胞培养。细胞护理和他们的适当 识别和选择是确保数据可重现的关键。评选 移植研究中使用的适当工具，如细胞的实时荧光标记 决定了这类研究的长期结果。将向调查人员提供关于 移植研究的设计和移植细胞的特征。 斑马鱼已经成为一种重要的脊椎动物模型，不仅用于研究 不仅用于研究神经发育，也用于研究行为、神经退行性变、代谢和其他 疾病过程，包括癌症。发展性研究尤其利用了这一点。 模型，因为胚胎的透明性，使得能够可视化特定的种群 神经元及其轴突使用绿色荧光标记的转基因，并有能力做高 吞吐量基因筛查。此外，斑马鱼转录本的序列和 基因组已接近完成。一大组影响神经系统发育的突变体 已经可以使用了。因此，人们对斑马鱼模型有很高的兴趣 MRRC的成员以及加州大学洛杉矶分校的整个生物医学研究社区。事实上, 加州大学洛杉矶分校生命科学大楼内的一个大型集中式斑马鱼设施目前正在 计划。需要这样的设施来容纳加州大学洛杉矶分校的调查人员，他们目前使用或 预计将使用这种模式，并用于招聘新的教师。MRRC热情地 支持这一提议，应能满足大屏幕的大部分空间需求 持有主要斑马鱼种群。然而，许多试验性工作将需要 要么在单独的实验室完成，要么在每栋建筑的公共设施中更高效地完成 由使用该模型的调查人员共享。公共设施的使用将会多得多 节省成本和时间，因为它将避免每个实验室都需要学习 方法和提供常规的养鱼，建立养殖等。它还将腾出空间在 用于其他项目的单独实验室。为此，我们专门为此设立了一个房间。 在新的神经科学研究大楼里，将容纳大多数MRRC调查人员。这个房间 大约350英尺2英寸，能够容纳3-5个鱼架。这间客房将由 作者：James A.Waschek；作者：Paul赵，SRA。Waschek医生最近回来了 从9个月的实验室休假或斯蒂芬·埃克博士在加州大学 他已经使用斑马鱼模型好几年了。在瓦谢克的监督下， 技术人员将维护所有鱼线，设置养殖，并为个体提供胚胎 调查人员。研究助理还将向个人传授胚胎注射技术 调查人员。

Purpose and Objectives Genetic and environmentally-induced developmental disorders can adversely affect central nervous system (CMS) development and function. The mechanisms of pathogenesis and neural repair as well as the translational research that will lead to the development of therapeutic treatments for regeneration of the damaged CNS have been increasingly the subject of intense study by researchers, including the members of this Center. To match the complexity of these problems one needs a multidisciplinary approach. Thus, it was to be useful to our neuroscientists to come to a single integrated neuroscience core where they could be advised by a team of expert faculty and staff of the proper approach, technique and equipment to use, to investigate their neuroscience research problems. To achieve this objective 3 cores were merged (Cell Biology, Cytology and Imaging). Expensive state-of-the-art instruments like the Zeiss Laser Scanning Confocal Microscope LSM 510 META, and the Leica Laser Cell Capture Microdissection system were purchased. Additional faculty were recruited to offer broad neuroscience expertise which together with contributions provided by other MRRC and Campus Cores, would satisfactorily address the vast needs of projects listed in the cores user table. Here the rationale for the services provided will be presented. The Neuroscience and Imaging Core provides expertise, services and equipment to support cellular and molecular neuroscience research: ¿ Stem Cell/ Cell Culture/Cell Transplant ¿ Zebrafish Facility ¿ Neurocytology ¿ Immunocytochemistry ¿ In Situ Hybridization ¿ Confocal Microscopy (Zeiss 510 META) ¿ Laser Cell Capture from tissue slices and cell cultures First, the in vitro/in vivo approaches of cell culture and cell transplantation will be discussed. The discovery that the adult CNS possesses the potential to regenerate via multipotent stem cells, has given rise to increasing interest on studying neural stem cells either to understand pathogenesis in neurodegenerative diseases in animal models or as potential therapeutic tools for degenerative diseases. The purpose of this core is to fulfill the need of incorporating new technology aimed at the study of the restoration of CNS function. The Core combines various specialized domains, including the culture of specific neural cells, neural stem and ES cells, their propagation, proper differentiation, identification and their use for neural transplantation. These approaches have proven to be promising in the field of CNS repair. The core integrates the state-of-the-art methodology on the preparation of embryonic stem cells (ES) and neural stem cells, as well as neuronal and glial cell cultures. Cell care and their proper identification and selection are of the essence to insure reproducible data. The selection of adequate tools such as live fluorescent labeling of cells to be used in transplant studies determines the long-term outcome of this type of studies. Investigators will be advised on the design of transplant studies and characterization of grafted cells. Zebrafish have become an important vertebrate animal model, not only for investigation of neural development but also for studies of behavior, neurodegenerative, metabolic and other disease processes, including cancer. Developmental studies in particular capitalize on this model, because of the transparency of embryos, enabling visualization of specific populations of neurons and their axons using green fluorescence-tagged transgenes, and the ability to do high throughput genetic screens. Moreover, the sequencings of zebrafish transcriptomes and the genome are near completion. A large set of mutants that affect nervous system development are already available. As such, there is a high level of interest in the Zebrafish model by members of the MRRC, as well as the overall biomedical research community at UCLA. In fact, a large centralized zebrafish facility in the Life Sciences building at UCLA is currently in the planning. Such a facility is needed to accommodate UCLA investigators who currently use, or expect to use this model, and for the recruitment of new faculty. The MRRC has enthusiastically supported this proposal, which should handle much of the space needs for large screens and the holding of major zebrafish stocks. However, much of the experimental work will need to be done either in individual laboratories, or more efficiently, in a common facility in each building shared by the investigators who use the model. Use of a common facility will be much more cost- and time-efficient because it will circumvent the need for each laboratory to learn the methods and provide routine care of fish, set up breeding, etc. It will also free up space in individual laboratories for other projects. To this end, we have dedicated a room for this purpose in the new Neuroscience Research building that will house most MRRC investigators. This room is approximately 350 ft2, and is capable of holding 3-5 racks of fish. This room will be managed by Dr. James A. Waschek with the help of Paul Zhao, SRA. Dr. Waschek recently returned from nine months of sabbatical leave at the laboratory or Dr. Stephen Ekker, at the University of Minnesota, who has used the zebrafish model for several years. Under Waschek's supervision, the technician will maintain all lines of fish, set up breedings, and provide embryos for individual investigators. The research associate will also teach embryo injection techniques to individual investigators.