CORE--NEUROCYTOLOGY /CELLULAR IMAGING

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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.