MORPHOLOGY AND CELL IMAGING CORE

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The Morphology and Cell Imaging Core has been providing expertise, technical support and assistance for morphological and imaging approaches as well as state-of-the-art equipment for sophisticated imaging techniques to the DDRCC members for over 18 years under the leadership of Dr. Sternini. The services provided by the core have been instrumental for studies that have elucidated the tissue and cellular distribution of signaling molecules that play a role in the control of gastrointestinal (Gl) function and in the pathogenesis of Gl disorders as well as for visualizing signaling pathways that regulate cellular functions. The studies that have benefited from the core have made important advances in our understanding of functional interactions between neuronal and hormonal pathways regulating Gl functions and have resulted in many publications and in long-lasting collaborations. There have been several additions and updates in services throughout the years to reflect the needs of DDRCC members and the evolving of morphological and imaging technologies, with the goal of enhancing the Core capabilities. Enhancement of morphological capabilities includes 1) the addition of mouse pathology services and 2) the incorporation of a Central Bank of well characterized antibodies. The mouse pathology services have been added in our previous competitive renewal in response to the increasing need of many CURE: DDRCC investigators for expertise and assistance in defining phenotypical abnormalities or pathologies that can be detected in genetically manipulated mice, which are increasingly used at CURE:DDRCC. Dr. Nora Rozengurt, an experienced veterinary pathologist with extensive expertise in natural and experimentally induced mouse pathology has been serving as Core Associate Director to supervise this service, which has been extensively used during the past funded cycle. The antibodies central bank has been added to provide a centralized system for the wealth of antibodies that have been produced at CURE during previous funding periods and for other antibodies that are used by many investigators as markers for specific cell types or intracellular signaling molecules. The availability of a centralized bank of well characterized and validated antibodies of interest to CURE: DDRCC investigators is a powerful resource that will allow an effective access of CURE:DDRCC investigators to a variety of reagents that can be utilized through the Morphology and Cell Imaging Core. Dr. Gordon Ohning will join the Core as Associate Director to be in charge of the maintenance of the Central Bank of antibodies. There has been a significant enhancement of the capabilities of our imaging services, including 1) the addition of highly sophisticated equipment and 2) the incorporation of new imaging technologies that will further advance our ability to visualize molecules at the cellular and subcellular levels and changes in second messenger systems following cellular activation. A major enrichment of our imaging facilities has been the addition of a state-of-the-art confocal microscope, a Zeiss LSM 510 Meta equipped with sophisticated software (e.g. deconvolution, image analysis, 3-D rendering) for live and fixed cell imaging. This system was acquired with the help of institutional funds from the David Geffen UCLA School of Medicine just prior to the previous competitive renewal, and since then it has been used heavily by an increasing number of CURE: DDRCC investigators. During the past funded cycle, we have also enhanced our instrumentation for calcium imaging by acquiring a Zeiss Pascal confocal microscope equipped with both short (405 nm) and visible wavelength lasers for excitation. A whole cell patch clamp recording setup is available to perform single cell electrophysiology simultaneously with confocal live cell imaging. This microscope, which has also been acquired with the help of institutional funds from the School of Medicine Dean's office, is housed at UCLA in the same building as the Zeiss 510 Meta confocal and has been extensively used by severa DDRCC investigators. Our services have also been upgraded by the addition of new imaging technologies in live cells, including quantum dots (Q-dots), photoactivatable proteins and second messengers' biosensors labeled with fluorescent tags. These significant improvements will further advance our ability to visualize macromolecules at the cellular and subcellular levels and changes in second messenger systems following cellular activation. Dr. Osvaldo Rey will join the Core personnel as Core Co-Director to supervise the new imaging methodologies that are proposed to cover the new needs of many DDRCC investigators. Finally, there will be an increased use of shared resources that are available for CURE: DDRCC investigators through arrangements made by the Core Director and existing Cores at UCLA, which are supported by the Department of Pathology and Laboratory Medicine, the Brain Research Institute and the Nanotechnology Institute. These interactions will continue to be a major effort of the Morphology and Cell Imaging Core to enhance the technical capabilities of the Core and ensure optimal utilization of shared resources within the UCLA community that includes the CURE: DDRCC community. Overall, the addition of services briefly outlined above and explained in more detail later in the body of the application reflects the evolving of the DDRCC research interests and focus, and the advances in morphological and imaging technologies. These additions will provide state-of-the-art approaches and equipment to many CURE: DDRCC investigators who are interested in elucidating fundamental normal or abnormal mechanisms in different types of cells controlling digestive functions.