Robert E. Marc - US grants

The long term goals of this research are to: (1) generate a complete network map for the mammalian retina and (2) define the signaling profiles of all retina cells. Fusions of molecular and computational analysesnow make it possible to perform these tasks in a practical time frame. Specific Aim 1. Realization of a complete network map for the mammalian retina. Retinal networks converge on ganglion cells, creating some 15-20 filtered versions of the visual world. Maps of these filters will be acquired by fusing molecular phenotyping (to visualize all cells), excitation mapping (to visualize function), and large-scale ultrastructural imaging (to visualize all connections). The novelstrategy is the computational propagation of molecular data into the ultrastructural data, producing a combined map of all cell classes and connections. Significance: Understanding any system requires a complete map: a map against which changes triggered by disease or experimental intervention can be gauged. This work has taken on new importance as inherited or acquired retinal degenerations are now known to heavily impact retinal wiring and neuronal survival, showing significant similarities to temporal lobe epilepsy. A complete network map provides the essential guide for any strategy to restore vision. Specific Aim 2. Definition of the basic signaling profiles of all retinal neurons. Each of the60+ retinal neuron classes expresses a profile of molecular mechanisms that precisely tunes it for vision. These profiles will be acquired for all neuronal classes by fusing molecular phenotyping and visualization of function with organic-cation permeation and D-glucosamine metabolic mapping. The glucosamine method is based on the same metabollic principles as fMRI brain imaging, linking cellular and systems visualizations. Permation mapping reports integrated cell currents while metabolic mapping reports integrated cell voltages and each provides a subtly different view of signaling. Significance: A description of any network requires a functional profile for each cell. These profiles shape the visual performances of ranges of each cell. We now know now that these profiles are not static; that they are responsive to developmental experience and are altered by retinal degenerations. Any strategies to restore vision must preserve or restore to the molecular profiles of the key pathways defined in Specific Aim 1.

biomedical equipment resource; biomedical equipment purchase;

DESCRIPTION (provided by applicant): A cascade of neuronal and glial transformations are triggered by photoreceptor or retinal pigmented epithelium degenerations. These remodeling processes, initiated prior to the death of photoreceptors and persisting after they are lost, encompass over 30 distinct aberrations including altered glutamate receptor expression, anomalous neurite and synapse formation, anomalous signaling, intraretinal migration and emigration, and neuronal death. Remodeling stands in the way of many proposed therapies. Our program focuses on discovering key remodeling mechanisms for drug discovery, intervention and retinal stabilization in retinitis pigmentosa (RP) and AMD. We will attempt this through three aims: (1) analysis of early cone and bipolar cell deconstruction;(2) exploration of mechanisms of corruptive intrinsic non-visual self-signaling and retinoid-induced axonogenesis;and (3) characterizing remodeling in AMD / AMD-like retinal degenerations. This research fuses advanced cell profiling methods of excitation mapping, computational molecular phenotyping (CMP), and traditional protein mapping with a selection of RP/AMD animal models (guanylate cyclase knockout mice, light-induced retinal degeneration mice, Pde6brd1 mice, rhodopsin P347L transgenic rabbits). We seek definition of key molecular pathologies (e.g. MAPK pathways in cones and bipolar cells, aberrant retinoid processing;emigrant cell profiles) that may serve as drug targets. The goal is to preserve RP and AMD retinas for genetic, molecular, cellular or bionic rescue. Relevance. Loss of neuronal structure and function in remodeling is likely the earliest cause of vision loss in retinal degenerations. These anomalies progressively impede development of therapies. Discovering the mechanisms of the many forms of remodeling may facilitate neuroprotective drug development or identification of existing drugs. PUBLIC HEALTH RELEVANCE: Loss of neuronal structure and function in remodeling is likely the earliest cause of vision loss in retinal degenerations. These anomalies progressively impede development of therapies. Discovering the mechanisms of the many forms of remodeling may facilitate neuroprotective drug development or identification of existing drugs.

DESCRIPTION (provided by applicant): Vision research at the University of Utah engages over 17 faculty members and 50+ supporting staff, graduate students and postdoctoral fellows. This proposal for a Vision Research Core Grant will enhance the programs of twelve participating investigators who collectively hold fifteen R01 awards from the National Eye Institute. The Core consists of (1) Imaging, (2) Functional Assessment, and (3) Molecular Biology modules, providing comprehensive resources, training and technical assistance to the programs of the participating investigators. In addition, a novel client-server application (iLab) will provide secure, uniform workflow and data management for all modules, facilitating access, conflict resolution, job scheduling and data tracking. This tool assures data integrity and any dataset can be collaboratively accessed. The Imaging Module will provide: (1) high-resolution optical imaging with high-speed image capture; (2) laser confocal scanning microscopy (LSCM); (3) electron microscopy (EM) with high-resolution digital capture; (4) computational fusion of optical, LSCM, and EM data; (5) ultramicrotomy cryomicrotomy services, including immunocytochemistry; (6) training; (7) resource maintenance; and (8) comprehensive data archiving. The Functional Assessment Module will provide: (1) comprehensive ERG/VEP visual testing; (2) optomotor behavioral tracking; (3) optical imaging of intrinsic CMS signals; (4) optical system calibration; (5) training; (6) resource maintenance; and (8) comprehensive data archiving. The Molecular Biology Module will provide: (1) high-capacity gene sequencing (2) robotics for sample processing; (3) IR imaging of blots and gels; (4) high-capacity ISH screening (5) training; (6) resource maintenance; and (7) comprehensive data archiving. Access to new technologies, training and technical support will facilitate collaborations, allow participating investigators to explore new research avenues and funding opportunities, and enhance the educational experience of graduate students, medical students, interns, residents and postdoctoral fellows.

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ABSTRACT NOT PROVIDED

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The long term goals of this research are to: (1) generate a complete network map for the mammalian retina and (2) define the signaling profiles of all retinal cells. Fusions of molecular and computational analyses now make it possible to perform these tasks in a practical time frame. Specific Aim 1. Realization of a complete network map for the mammalian retina. Retinal networks converge on ganglion cells, creating some 15-20 filtered versions of the visual world. Maps of these filters will be acquired by fusing molecular phenotyping (to visualize all cells), excitation mapping (to visualize function), and large-scale ultrastructural imaging (to visualize all connections). The novel strategy is the computational propagation of molecular data into the ultrastructural data, producing a combined map of all cell classes and connections. Significance: Understanding any system requires a complete map: a map against which changes triggered by disease or experimental intervention can be gauged. This work has taken on new importance as inherited or acquired retinal degenerations are now known to heavily impact retinal wiring and neuronal survival, showing significant similarities to temporal lobe epilepsy. A complete network map provides the essential guide for any strategy to restore vision.

DESCRIPTION (provided by applicant): The overall aims of this Vision Research Core (VRC) are to provide: access to resources outside the scope of individual R01 awards, access to technical expertise outside the scope a single laboratory, staff training to remove barriers to efficient translational research and collaboration. The research areas supported by the VRC span the analysis and treatments of retinal degenerations, developmental disorders, glaucoma and other disorders, as well as and a range of cutting-edge basic science initiatives. We have implemented four resource modules that continue the natural evolution of how this research group works together, serving 15 investigators holding 21 NEI R01 awards. The Imaging Module provides a range of imaging (TEM, confocal, scanning optical) and computing services (imaging, database, mathematics) based on strengths of core laboratories and the tradition of excellence of the UU School of Computing, whose descendants founded Adobe Systems, Silicon Graphics, Netscape and Pixar. Collaborations among these groups have transformed software tools for TEM and confocal imaging. The Histomics Module derives from the strengths in mouse genetics at the Moran Eye Center and the high demand for tissue profiling. Almost every JMEC laboratory has need of high quality histomics (cryosectioning, paratoming, genotyping, immunocytochemistry, PCR, ISH). The Physiology Module is a powerful set of tools that VRC faculty use for animal model validation and disease profiling: ERGs, OptoMotry behavioral testing, and a range of ocular imaging tools (OCT, Micron). Our new module includes in vivo/in vitro 2-photon imaging. The Biochemistry Module evolved from a gene sequencing service towards a true proteomics (ultracen- trifugation, gel quantitation) and metabolomics resource (specialized GC-MS, HPLC-MS, surface plas- mon resonance) more accurately reflecting our current research strengths. The Administrative Module provides professional grants management for the VRC.

DESCRIPTION (provided by applicant): Vision research now spans basic science (biology, chemistry, computer science, genetics, optics) but also has strong traditional links to neuroscience and newly evolving aspects of translational research in ocular disease. The principle underlying graduate education at the University of Utah is that students learn basic research and applied vision science, and experience strong interdisciplinary, interdepartmental collaborations. Predoctoral and postdoctoral trainees are endowed with skills ranging over molecular biology, electrophysiology, developmental neuroscience, retinal connectomics, translational science and visual behavior. These diverse approaches are unified by training in two intellectual streams that represent the core strengths of the Moran Vision Institute and the vision research community at the University of Utah: the study of human disease, a tradition of excellence in the cutting-edge praxis molecular biology, physiology and connectomics. Eighteen NIH/NEI-funded training faculty members are aligned in five broad areas of vision research: Molecular and Cellular Science, Developmental Science, Ocular Disease, Translational Interventions, and Computational Science. These areas reflect the breadth of the program and provide exceptional opportunities to the 74 current predoctoral and postdoctoral trainees. This application requests funding for 2 predoctoral and 4 postdoctoral trainees. Due to the outstanding, accredited predoctoral training programs already in place, recent graduates University of Utah have been placed in excellent postdoctoral fellowships at Yale, Stanford, Harvard, U Mass, UCSD, and UCSF. Our postdoctoral trainees have published widely, taken faculty positions, received Research to Prevent Blindness Career Development Awards, and are competing successfully for NIH RO1 funding. We believe that trainees from our Vision Research Training Program will become future leaders in vision research.

Project Summary The OVERALL aims of this Vision Research Core (VRC) are to provide: · access to resources outside the scope of individual R01 awards · access to technical expertise outside the scope a single laboratory · staff training to remove barriers to efficient translational research and collaboration The research areas supported by the VRC span the analysis and treatments of retinal degenerations, developmental disorders, glaucoma and other disorders, as well as and a range of cutting-edge basic science initiatives. We have implemented four resource modules that continue the natural evolution of how this research group works together, serving 15 investigators holding 21 NEI R01 awards. The Administrative Module provides professional grants management for the VRC. Specifically the Administrative Module provides: · 0.25 FTE services of Julee LaMothe (Sponsored Projects Professional) · Post-award grants management services · VRC ordering, supply tracking, recharge monitoring, resource scheduling, tracking equipment service agreements, scheduling service and repair · Assistance for the administrative activities Module Directors.

Project Summary The OVERALL aims of this Vision Research Core (VRC) are to provide: · access to resources outside the scope of individual R01 awards · access to technical expertise outside the scope a single laboratory · staff training to remove barriers to efficient translational research and collaboration The research areas supported by the VRC span the analysis and treatments of retinal degenerations, developmental disorders, glaucoma and other disorders, as well as and a range of cutting-edge basic science initiatives. We have implemented four resource modules that continue the natural evolution of how this research group works together, serving 15 investigators holding 21 NEI R01 awards. The Imaging Module provides a range of imaging (TEM, confocal, scanning optical) and computing services (imaging, database, mathematics) based on strengths of core laboratories and the tradition of excellence of the UU School of Computing, whose descendants founded Adobe Systems, Silicon Graphics, Netscape and Pixar. Collaborations among these groups have transformed software tools for TEM and confocal imaging. Specifically, the Imaging Module provides: · novel, powerful high-speed automated TEM for VRC investigators, regardless of experience, that de- livers top quality TEM imagery in a readily navigable format using the Viking web-application. · research grade confocal resources and management both in the JMEC proper and the JMEC vivar- ium; expanded confocal resources are planned. · high-speed optical scanning microscopy for high-throughput tissue / immunocytochemical analysis · extensive imaging, database and mathematics software holdings and training expertise · large scale data storage for the VRC (up to 0.5 petabyte)